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Everyone, come to look into laboratory cell culture technology to reveal the secret
when it comes to cell culture technology, you may not be unfamiliar with it. It runs through all aspects of cell detection experiments and is the basic factor that determines the success or failure of the experiment. Cell culture refers to a culture technique in which cells are removed from tissues in the body to mimic the environment in the body, under sterile, appropriate temperature, pH and certain nutritional conditions, to grow and reproduce, and to maintain their structure and function. It is used in cytology, Play an important role in the research and application of genetics, virology, and immunology. Cell culture is also a basic technology for virus research and vaccine development. Therefore, cell culture technology has been widely used in genetics, immunology, oncology, virology, molecular biology and other fields. Nuclear transfer, cell hybridization, DNA-mediated gene transfer and the establishment of some physical maps developed in recent years also need to be closely integrated with cell
How many scientific researchers have been frustrated because of the poor state of cell culture! How many laboratories have ever had to terminate their carefully developed long-term experiments due to microbial contamination! So how should cells be cultured to keep them in a normal state? How can we avoid contamination of cultured cells?
Below, Abbkine cell culture method gives you the answer.
The general process of cell culture mainly includes the following points:
Preparatory work: The content of the preparatory work includes the cleaning, drying and disinfection of utensils, the preparation, distribution and sterilization of culture medium and other reagents, the cleaning and disinfection of sterile rooms or ultra-clean benches, and the preparation of incubators and other instruments. Check and debug.
Extracting materials: Take out certain tissue cells from the body in a sterile environment (depending on the purpose of the experiment), and put them into a culture vessel after certain treatments (such as digestion and dispersion of cells, separation, etc.). This process is called extraction . In the case of cell line expansion, there is no such process as obtaining materials. The first culture of tissue cells taken out of the body is called primary culture.
Cultivation: The process of putting the obtained tissue cells into a culture flask or culture plate is called culture. If it is a tissue block culture, connect the tissue block directly to the bottom of the culture vessel. After a few hours, the tissue block can be attached to the bottom, and then add the culture medium.
Cryopreservation and resuscitation: In order to preserve cells, especially mutant cells or cell lines that are not easily available, cells should be frozen. The freezing temperature is generally -196℃ of liquid nitrogen. Collect the cells into the cryopreservation tube and add a medium containing a protective agent (usually dimethyl sulfoxide or glycerol), freeze at a certain cooling rate, and finally save In liquid nitrogen. At extremely low temperatures, the storage time of cells is almost unlimited. The resuscitation generally adopts a quick-thaw method, that is, after removing the cryotube from the liquid nitrogen, immediately put it in 37°C water to make it melt quickly within one minute. Then transfer the cells to a culture vessel for culture.
The basic steps of cell primary culture:
Primary isolation cell culture refers to the separation of tissues from the donor body into single cells or monotype cell groups by mechanical and digestion, so that they can simulate the physiological environment of the human body in vitro, under sterile, appropriate temperature and certain nutritional conditions. Survive, grow and reproduce. Primary cultured cells often have different cell components and grow slowly, but they are more representative of the tissue cell type from which they are derived and the specific characteristics of the expression tissue. Using primary cell culture to do various experiments, such as drug testing, cell differentiation and virology, has a good effect. The steps are as follows:
Cut the tissue: first clean the obtained tissue with D-Hanks or Hanks solution to remove blood stains on the surface, and use surgical forceps to remove the attached connective tissue and other non-cultivating tissues. After cleaning again, use a scalpel to cut the tissue into several small pieces, transfer them into a penicillin vial or a small beaker, add an appropriate amount of buffer, and use elbow ophthalmic scissors to repeatedly cut the tissue until the tissue becomes a paste, about 1mm3 in size. After standing for a while, use a straw to suck off the upper layer of liquid, add appropriate buffer solution and wash again.
Digestion and separation: The purpose of digestion and separation is to digest and separate small tissue pieces into cell clusters or scattered single cells to facilitate further culture. Commonly used digestive enzymes are trypsin and collagenase.
Cultivation: Use a counting plate to count the cells of the cell suspension. Adjust the number of cells to (2～5)×105 cells/ml with the culture medium, or the density required for the experiment, and divide them into culture flasks so that the amount of cell suspension is slightly higher than the bottom of the culture flask after covering. Place in a CO2 incubator, 5% CO2, 37°C static culture. Generally 3~5d, the primary cultured cells can adhere to the wall of the bottle and stretch and start to grow. You can add a new medium of 1/2 of the original medium, continue to culture for 2~3d and then change the medium, generally 7~14d can grow Fill the wall of the bottle and pass it down.
Subculture
Precautions
Digestion time: Master the time for cell digestion. When the digestion time is too short, the cells should not fall off the bottle wall. Excessive digestion will cause the cells to fall off and damage.
Digestion concentration: master the digestion concentration. When the digestion solution concentration is too high, the digestion time should be shortened, and when the digestion solution concentration is too low, the cell digestion time will be relatively prolonged.
Significance of cell cryopreservation
It is a technology that puts cells in a low temperature environment (liquid nitrogen) to reduce cell metabolism for long-term storage. Cryopreservation of cells is one of the main methods of cell preservation, which plays a role in cell preservation.
Significance of cell recovery
Cell resuscitation, a term in biology, refers to the process of re-cultivating cells frozen in liquid nitrogen or in a refrigerator at -70°C to restore growth.
For the great love of “Yinfan”, we will have great scientific research surprises waiting for everyone. At the same time, in order to provide a platform for readers to learn biological knowledge, Abbkine’s technical experts have worked tirelessly to sort out the dry goods of a variety of high-end biological experiments, and later published them in the WeChat public account. Please continue to pay attention to Abbkine for readers who love science the public.
Abbkine focuses on the fields of proteinology and cytology, and is committed to the innovation and research and development of various antibodies, proteins, analytical reagents and kits, in order to become a key promoter in the development of life science research, drug development and other fields. We provide you with the favorite products of protein and immune research users, from basic immunological products, such as protein extraction and quantification, to internal reference label antibodies, primary antibodies and secondary antibodies for immunological experiments; the favorite products of cell research users, from Dyes and kits for detecting cell status, organelle extraction kits, cell substructure staining and tracking and cell metabolism detection products, to cytokine and protein detection kits for cell culture, just to help your research career !
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Abbkine LDH test kit, we want to say...
The last issue of Abbkine’s WeChat public account article tells about an important indicator of cell state detection, witnessing the state of cell viability. This is also the “good news” for many cytology researchers. Once the article was published, the response was very good, and everyone asked questions:
A: The Cell Living Dead Kit is so easy to use, we also want to start it. What is its purchase link?
B: What is the quotation for this kit? Is it family price?
C: Abbkine’s Living Dead Staining Kit is so easy to use, is there any other kit that can be used together?
D: In our laboratory, we do cell toxicology experiments. In addition to testing the effects of drugs on cell viability, can we also detect other states of cells?
For product purchase links and price questions, please leave a message below the article, and Abbkine’s beautiful little brothers and sisters will enthusiastically answer for everyone. For the combination of products, Abbkine, in order to help you detect more cell status indicators and more comprehensively evaluate the impact of external stimuli on cells, we have launched the LDH kit cytotoxicity detection kit (KTA1030).
Product NO. Product Name Product Links KTA1030 LDH Cytotoxicity Assay Kit https://www.abbkine.com/product/ldh-cytotoxicity-assay-kit-kta1030/
Popular science: lactate dehydrogenase (LDH or LD) is an important enzyme that catalyzes the redox reaction between lactic acid and pyruvate in glycolysis and gluconeogenesis engineering. Lactate dehydrogenase exists in the cytoplasm of all tissues and cells of the body, among which the content of the kidney is higher. In terms of the rate of glycolysis, lactate dehydrogenase is not a rate-limiting enzyme, so it has little effect on the rate of occurrence. Using this material, we quantitatively evaluate cell death or cytotoxicity through plasma membrane damage. LDH is a stable enzyme that exists in all cell types and is quickly released into the cell culture medium after the plasma membrane is damaged. Therefore, LDH is the most widely used marker in cytotoxicity studies.
The application range is very wide:
Analyze the effects of compounds or environmental factors on cell death. All animal cell samples can be tested. Detection range of cell number: 10,000-100,0000 cells/well.
Easy to operate:
No need to draw a standard curve, use positive and negative results to quantify cytotoxicity;
LDH is released from the cells into the medium, forming a visual color response. After the sample reaction is colored, use a microplate reader for detection, and the reading is convenient;
Samples and reaction equipment are easily available: only 200 μL of cell culture supernatant is required as a sample for subsequent testing. Ordinary 96-well plates can be completed.
This issue mainly talks about the experimental protocols and products of cytotoxicity detection in cytology research. Due to space limitations, I have to say goodbye to everyone again. At the same time in the next issue, there will be experiments and product application operation tips to meet with you. Stay tuned~~~~~
You are welcome to pay more attention to Abbkine’s official account, and there will be more scientific research surprises waiting for you. At the same time, in order to provide a platform for readers to learn biological knowledge, Abbkine’s technical experts have worked tirelessly to sort out the dry goods of a variety of high-end biological experiments, and later published them in the WeChat public account. Please continue to pay attention to Abbkine for readers who love science. the public.
Abbkine focuses on the fields of proteinology and cytology, and is committed to the innovation and research and development of various antibodies, proteins, analytical reagents and kits, in order to become a key promoter in the development of life science research, drug development and other fields. We provide you with the favorite products of protein and immune research users, from basic immunological products, such as protein extraction and quantification, to internal reference label antibodies, primary antibodies and secondary antibodies for immunological experiments; the favorite products of cell research users, from Dyes and kits for detecting cell status, organelle extraction kits, cell substructure staining and tracking and cell metabolism detection products, to cytokine and protein detection kits for cell culture, just to help your research career
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How to comprehensively study cell proliferation, the Abbkine EdU test kit turns a stone into gold
Cell proliferation is an important life characteristic of organisms, and cells proliferate by dividing. Single-celled organisms that produce new individuals through cell division. Multicellular organisms produce new cells through cell division to supplement aging or dead cells in the body. Multicellular organisms can develop into a new multicellular individual from a fertilized egg through cell division and differentiation. It must be emphasized that through cell division, the replicated genetic material can be evenly distributed to two daughter cells. It can be seen that cell proliferation is the basis of organism growth, development, reproduction and heredity.
It can be said that cell proliferation is accompanied by all the life processes of multicellular organisms. More and more researchers have focused on cell proliferation detection, and the methods for cell proliferation detection are constantly updated. So what are the methods to study cell proliferation so far?
Cell proliferation testing is a basic experimental method to evaluate cell health, genotoxicity and the effect of anti-tumor drugs. The most accurate method for detecting cell proliferation is the BrdU method. The EdU method detection kit is a revolutionary breakthrough in the Brdu method. EdU (5-bromo-2-deoxyuracil) kit is a pyrimidine analog that can be integrated into DNA double strands during DNA synthesis. Through the comparison of the above schematic diagrams, it is not difficult to find that the EdU method is a simple, reliable, and trouble-free innovative method for detecting cell proliferation without antibodies, no denaturation steps, and maintaining cell morphology and DNA integrity.
However, there are still many researchers who still do not get the most effective method for cell proliferation testing and save reaction time. At this time, Abbkine’s EdU test kit came into being with the joint efforts of R&D experts, which solved everyone’s urgent need. And Abbkine’s EdU detection kit has two models, which are matched with two different fluorescent channels to meet the needs of different customers for fluorescent channels. Increase the diversity of cell proliferation test results.
Product NO. Product NO. Fluorescence channel KTA2030 Cell Proliferation EdU Image Kit (Green Fluorescence) 488 channel KTA2031 Cell Proliferation EdU Image Kit (Orange Fluorescence) 549 channel
Moreover, the kit can not only label proliferating cells, but also use nuclear dyes to label the nuclei of all cells (including proliferating cells and non-proliferating cells). Proliferating cells can be obtained by the number of proliferating cells and the number of all cells. The ratio. The calculation result is accurate and reliable, and the application value is extremely wide. Here are some pictures of the test results of the Abbkine EdU test kit:
DAPI Channel
549 Channel
488 Channel
This issue mainly talks about the overall solution for cytology research. I have to say goodbye to everyone again. At the same time in the next issue, there will be experiments and product application operation tips to meet with you. Stay tuned~~~~~
You are welcome to pay more attention to Abbkine’s official account, and there will be more scientific research surprises waiting for everyone. At the same time, in order to provide readers with a platform for learning biological knowledge, Abbkine’s technical experts have worked tirelessly to sort out the dry goods of a variety of high-end biological experiments, which will be published in the WeChat public account in the future. Please continue to pay attention to Abbkine for readers who love science. the public.
Abbkine focuses on the fields of proteinology and cytology, and is committed to the innovation and research and development of various antibodies, proteins, analytical reagents and kits, in order to become a key promoter in the development of life science research, drug development and other fields. We provide you with the favorite products of protein and immune research users, from basic immunological products, such as protein extraction and quantification, to internal reference label antibodies, primary antibodies and secondary antibodies for immunological experiments; the favorite products of cell research users, from Dyes and kits for detecting cell status, organelle extraction kits, cell substructure staining and tracking and cell metabolism detection products, to cytokine and protein detection kits for cell culture, just to help your research career !
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How to choose flow cytometry antibody?
Flow cytometry is a very practical technique. It is characterized by convenience and quickness, accurate results, and successful sorting of target cells. However, when it comes to the specific selection of flow cytometry antibodies, it is estimated that many small partners will have difficulties. It is not so easy to obtain a beautiful flow cytometry result graph. The most important thing to consider before the experiment is the choice of antibody and the combination of fluorescein.
What issues should be paid attention to when selecting flow cytometry antibodies?
1.Basic requirements for antibodies
a.Know clearly whether the target to be tested is intracellular or extracellular protein;
b.Select antibody reactivity in strict accordance with the source of the sample;
c.You must choose antibodies that are clearly labeled for use in flow cytometry experiments and cannot be replaced by ordinary antibodies.
2.Requirements for flow cytometry
a.Understand that the flow cytometer used has several lasers, the common lasers are 405nm, 488nm and 633nm, etc. At present, most flow cytometers are equipped with 2 lasers of 488nm and 633nm;
b.Know how many detection channels are under each laser, and the number of channels determines how many indicators can be measured at a time;
Instrument Common Excitation （Ex） light Fluorescence channel Common fluorescent label flow cytometry 488nm 530/30 FITC, DyLight 488, Abfluor 488 575/26 PE 610/20 PE/TR 695/40 PerCP/Cyanine5.5、 PE/Cyanine5、PerCP 780/60 PE/Cyanine7 633nm 660/20 APC、 Abfluor 647 730/45 Abfluor 750 780/60 APC/Cyanine7
3.Choice of fluorescent label
a.If there is a direct-labeled antibody, try to choose the direct-labeled antibody as much as possible. Indirect labeling of the secondary antibody increases the experimental steps. Multiple washings will cause cell loss and low staining accuracy;
b.Weakly express the target, try to choose a strong fluorescein-labeled antibody, PE> APC> FITC> PerCP.
4.Multi-color matching scheme
a.Each channel can only use one type of label, such as FITC and AF488. The excitation and emission wavelengths of the two labels are similar, so these two labels cannot be used at the same time;
b.Minimize spectral overlap, such as PE-cy5 and APC, adjust the compensation value to be too large when both are used;
c.Weak targets are equipped with strong fluorescein. For example, the expression of CD4 in Treg cells is higher than FoxP3, so FoxP3 should use strong fluorescein-labeled antibodies;
d.It is recommended to consult the literature for five-color and above color matching, or consult the antibody manufacturer.
5.How to adjust compensation?
In multicolor flow experiments, due to the overlap of the spectra of each fluorescein, compensation adjustment is required. The following figure shows the spectra of FITC and PE fluoresceins. Near 550nm, the two dyes have spectral overlap.
6.Isotype control?
The purpose of isotype control is to reduce background interference and better display the binding strength of antigen and antibody. Especially for some intracellular proteins, low expression or continuous expression proteins, the setting of isotype control is particularly necessary. Isotype control needs to select antibodies with the same species source, the same antibody subtype, and the same fluorescent label as the stained monoclonal antibody.
Recommendation: Elabscience flow cytometry antibody one-a more cost-effective overall solution for flow cytometry experiments
Elabscience has selected clone numbers reported many times in the literature as the source of flow cytometry antibodies, anti-human/rat/mouse, etc., and has a wealth of indicators, with> 1480 online indicators covering most common flow cytometry antibodies, and There are 12 kinds of fluorescent labels, which can meet the needs of many kinds of experiments. In addition, all fluorescent antibodies provide convenient Test specifications for your direct use. Most importantly, Elabscience can also customize the flow color scheme for free, so you don’t have to worry about multicolor experiments.
Product example image:
Figure six indicators are detected simultaneously, the negative and positive cell populations of all indicators can be fully separated.
About Abbkine
Our position: to serve global users of cell and protein research, to provide users with economical and technical product solutions in the form of application processes and product portfolios.
Our mission: to stimulate our inner creativity, provide competitive biomedical products and services, and continue to create maximum value for customers.
Our vision: to be a respected, world-class supplier of biomedical products and services.
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High Uric Acid Level and Gout
Uric acid is a natural waste product from the digestion of foods that contain purines. Purines are normally produced in the body and are also found in some foods and drinks. Foods with high content of purines include liver, anchovies, mackerel, dried beans and peas, and beer.
Gout occurs when urate crystals accumulate in your joint, causing the inflammation and intense pain of a gout attack. Urate crystals can form when you have high levels of uric acid in your blood.
Your body produces uric acid when it breaks down purines — substances that are found naturally in your body.
Fig 1: Metabolic pathways of uric acid formation from nucleotide monophosphates.
(Picture Source: https://pubmed.ncbi.nlm.nih.gov/24287461/ )
Through year of efforts, Abbkine research scientists have developed a series of assay kits for cell metabolism and that constitutes Abbkine CheKine™group. For detailed index, you are welcome to view at https://www.abbkine.com/?s=CheKine%E2%84%A2&post_type=product&submit=. In which we can see the detection kits of NAD/NADH, NADP/NADPH, superoxide dismutases (SOD), peroxidase (HRP) and glucose, but today we are going to introduce the test of uric acid for you.
To more specific, the benefits of Abbkine Uric Acid (UA) Colorimetric Assay Kit can be concluded as below:
Determination of Uric Acid (UA) concentration in a variety of biological samples such as animal tissues, serum,urine and other biological fluids.
Detailed sample preparation and result calculation methods are provided.
Simple operation steps.
You can view the kit protocol at https://www.abbkine.com/file/booklet/KTB1510-B.pdf
Kit Components:
Extraction Buffer
Reagent I A
Reagent I B
Standard
Abbkine is aimed at to provide high quality and cost effective reagents to global scientists, this Uric Acid (UA) Colorimetric Assay Kit has been optimized to use and can deduct your experiment cost. For detailed product information, you can refer in below sheet.
CAT# Product Name Size and Price KTB1510 CheKine™ Uric Acid (UA) Colorimetric Assay Kit $90/48T
$160/96T
If you are interested in Abbkine CheKine™Uric Acid (UA) Colorimetric Assay Kit, feel free to contact [email protected], our service representative will get back to you shortly!
About Abbkine Scientific Co., Ltd.
Abbkine offers services for global scientists in the fields of proteomics and cytology, and is committed to the innovation and development of various scientific reagents related to proteomics and cytology, to promote the development of fields including life science research and drug discovery. Proteomics products cover the preparation of samples (protein extraction, purification, coupling), protein quantification, antibodies and kits for protein detection. Cytology products involve cytokines (cell culture), cell status detection, cell staining, organelle extraction, cell metabolism and cytopathology reagents (kits). Abbkine relies on the product portfolio and unique marketing support as the main market strategy and product innovation mode, to help your research career!
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New Product Arrival, ALT and AST Test Kit Available from Abbkine!
Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) are enzymes found mainly in the liver, but also found in red blood cells, heart cells, muscle tissue and other organs, such as the pancreas and kidneys. An AST test is often performed along with an alanine aminotransferase (ALT) test. Both are enzymes found in the liver that become elevated in the blood when the liver is damaged.
AST is usually done as part of a group of liver function tests called a liver panel. It’s often ordered with a test for alanine aminotransferase (ALT), another liver enzyme. ALT is more accurate than AST at detecting liver disease. Abbkine has newly launched ALT test kit to help scientific research.
Fig 1: TCA Tycle
(Picture Source: https://www.mdpi.com/1422-0067/20/2/252)
Through year of efforts, Abbkine research scientists have developed a series of assay kits for cell metabolism and that constitutes Abbkine CheKine™group. In which we can see the detection kits of NAD/NADH, NADP/NADPH, superoxide dismutases (SOD), peroxidase (HRP) and glucose, but today we are going to introduce the test of ALT for you.
To more specific, the benefits of Abbkine Alanine Aminotransferase (ALT/GPT) Activity Colorimetric Assay Kit can be concluded as below:
Determination of ALT/GPT activity in a variety of biological samples such as animal tissues, plant tissues, cell culture: adherent or suspension cells, serum, bacterial.
Detailed sample preparation and result calculation methods are provided.
Simple operation steps.
Standard and standard curve line are provided.
You can view the kit protocol at https://www.abbkine.com/file/booklet/KTB1410-B.pdf .
Kit Components:
Extraction Buffer
Reagent I
Reagent II
Reagent III
Standard
Abbkine is aimed at to provide high quality and cost effective reagents to global scientists, this Alanine Aminotransferase (ALT/GPT) Activity Colorimetric Assay Kit has been optimized to use and can deduct your experiment cost. For detailed product information, you can refer in below sheet.
CAT# Product Name Size and Price KTB1410 CheKine™ Alanine Aminotransferase (ALT/GPT) Activity Colorimetric Assay Kit $40/96T
If you are interested in Abbkine Alanine Aminotransferase (ALT/GPT) Activity Colorimetric Assay Kit, feel free to contact [email protected], our service representative will get back to you shortly!
About Abbkine Scientific Co., Ltd.
Abbkine offers services for global scientists in the fields of proteomics and cytology, and is committed to the innovation and development of various scientific reagents related to proteomics and cytology, to promote the development of fields including life science research and drug discovery. Proteomics products cover the preparation of samples (protein extraction, purification, coupling), protein quantification, antibodies and kits for protein detection. Cytology products involve cytokines (cell culture), cell status detection, cell staining, organelle extraction, cell metabolism and cytopathology reagents (kits). Abbkine relies on the product portfolio and unique marketing support as the main market strategy and product innovation mode, to help your research career!
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New Product Arrival, ALT and AST Test Kit Available from Abbkine!
Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) are enzymes found mainly in the liver, but also found in red blood cells, heart cells, muscle tissue and other organs, such as the pancreas and kidneys. An AST test is often performed along with an alanine aminotransferase (ALT) test. Both are enzymes found in the liver that become elevated in the blood when the liver is damaged.
AST is usually done as part of a group of liver function tests called a liver panel. It’s often ordered with a test for alanine aminotransferase (ALT), another liver enzyme. ALT is more accurate than AST at detecting liver disease. Abbkine has newly launched ALT test kit to help scientific research.
Fig 1: TCA Tycle
(Picture Source: https://www.mdpi.com/1422-0067/20/2/252)
Through year of efforts, Abbkine research scientists have developed a series of assay kits for cell metabolism and that constitutes Abbkine CheKine™group. In which we can see the detection kits of NAD/NADH, NADP/NADPH, superoxide dismutases (SOD), peroxidase (HRP) and glucose, but today we are going to introduce the test of ALT for you.
To more specific, the benefits of Abbkine Alanine Aminotransferase (ALT/GPT) Activity Colorimetric Assay Kit can be concluded as below:
Determination of ALT/GPT activity in a variety of biological samples such as animal tissues, plant tissues, cell culture: adherent or suspension cells, serum, bacterial.
Detailed sample preparation and result calculation methods are provided.
Simple operation steps.
Standard and standard curve line are provided.
You can view the kit protocol at https://www.abbkine.com/file/booklet/KTB1410-B.pdf .
Kit Components:
Extraction Buffer
Reagent I
Reagent II
Reagent III
Standard
Abbkine is aimed at to provide high quality and cost effective reagents to global scientists, this Alanine Aminotransferase (ALT/GPT) Activity Colorimetric Assay Kit has been optimized to use and can deduct your experiment cost. For detailed product information, you can refer in below sheet.
CAT# Product Name Size and Price KTB1410 CheKine™ Alanine Aminotransferase (ALT/GPT) Activity Colorimetric Assay Kit $40/96T
If you are interested in Abbkine Alanine Aminotransferase (ALT/GPT) Activity Colorimetric Assay Kit, feel free to contact [email protected], our service representative will get back to you shortly!
About Abbkine Scientific Co., Ltd.
Abbkine offers services for global scientists in the fields of proteomics and cytology, and is committed to the innovation and development of various scientific reagents related to proteomics and cytology, to promote the development of fields including life science research and drug discovery. Proteomics products cover the preparation of samples (protein extraction, purification, coupling), protein quantification, antibodies and kits for protein detection. Cytology products involve cytokines (cell culture), cell status detection, cell staining, organelle extraction, cell metabolism and cytopathology reagents (kits). Abbkine relies on the product portfolio and unique marketing support as the main market strategy and product innovation mode, to help your research career!
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How much do you know about tissue sections? Abbkine analyzes for you one by one
Not long ago, Yacoin organized a full-blown experimental study of immunopathology, which aroused the interest of many curious babies. Everyone left messages, and the pathology experiment also included immunohistochemistry, pathology detection and other aspects, and there was still a lot of pathology knowledge that was not shown to everyone. Seeing the questions you have left, the editor will announce them one by one on the Yacoin WeChat platform in the follow-up. In this issue, the editor will start from the shallower to the deeper, starting with the knowledge of tissue slices for everyone.
When it comes to tissue sectioning, you will have many questions. Here is an analysis of your questions one by one:
What is a tissue section?
A type of qie pian slide specimen. A thin slice of animal and plant tissues for optical or electron microscope observation. Due to different requirements, a blade can be used for freehand sectioning, or the tissue block can be embedded in paraffin or collodion or frozen at low temperature and sectioned with a microtome. Cut into 5~10 micron slices for observation by optical microscope. Ultra-thin sections cut with epoxy resin or methacrylic acid embedded tissue blocks, with a thickness of 20-50 nanometers, designed for observation under an electron microscope. The sections of root tips and stems used in general teaching are generally called paraffin sections.
What is the initial shape of the tissue specimen of the tissue section?
The size of the tissue block: the ideal volume of the tissue block is 2.0cm×2.0cm×0.3cm, so that the fixative can penetrate into the tissue quickly and evenly. However, the tissue block is ideal according to the different preparation materials and purposes The volume is also different. For example, when making pathological examinations and scientific research sections, the tissue block can be thinned by 0.1~0.2cm, which can shorten the time of fixation, dehydration and transparency. If the thickness of the teaching section is 0.3~0.5cm, the same wax can be used. Block makes more teaching slices.
How to make tissue dehydration transparent?
After the specimen is fixed and washed, the tissue contains a lot of water, and the water in the tissue block must be replaced. This process is called dehydration. Whether it is sliced with paraffin or collodion, the water in the tissue must be removed. Because water-containing tissue is incompatible with embedding materials such as paraffin wax and collodion, the commonly used dehydrating agent is a series of ethanol with different concentrations. .
The dehydration steps are: 80%, 90%, 95%, 100% ethanol of various concentrations for 2 hours, this process can be completed by the automatic control of the dehydrator.
Acetone is also a dehydrating agent with strong dehydration ability, but because of its strong dehydration ability, it has a violent contraction effect on tissues, so it is generally not used when making scientific research and teaching sections. Because ethanol, acetone, etc. are not soluble in paraffin, a solvent replacement process that can dissolve in paraffin is called transparent. Commonly used transparent agents are xylene, chloroform, methyl salicylate, etc.
How to perform wax immersion, embedding, sectioning, and patching?
(1) Trimming wax block: Depending on the size of the tissue, cut off the remaining wax at about 0.1~0.2cm at the edge of the tissue, otherwise the tissue may shrink and become uneven.
(2) Prepare slicing tools: slicing knife, writing brush, ophthalmic tweezers (curved), bleaching temperature controller
(3) Install wax block: install the repaired wax block on a metal or wooden wax holder.
(4) Install the slicing knife: Install the slicing knife on the knife table of the slicing machine, and tighten the fastening screws on the knife table to prevent vibration during slicing and maintain a certain slice thickness.
(5) The thickness of the slice: the thickness regulator of the slicer is engraved with 0~50μm or 0~25μm, and the thickness can be selected arbitrarily. The thickness of the paraffin slice is generally 4~6μm.
(6) Slice
(7) Spreading: Use ophthalmic tweezers to lift the wax tape and gently spread it on the water surface at 40~45°C. Use the tension of the water and the temperature of the water to naturally flatten the slightly wrinkled wax tape.
(8) Patches and baking sheets: After the slices are fully flattened on the constant temperature water surface, the wax slices are pulled to the middle of the slide glass and the remaining water on the slide glass is poured, and placed in a 60-65 ℃ incubator Bake the slices in the oven of the temperature controller for 15 to 30 minutes to remove the paraffin that melts the tissue gap.
The picture shows a set of experimental equipment for paraffin sectioning: wax embedding machine, slicer, and spreader.
The picture shows the standard operation method of paraffin section。
How to improve the level of tissue sectioning?
1) Pathology instrument is a necessary condition: choosing the right slicer is the primary factor of the experiment;
2) The quality of the sample to be cut is a key factor: the quality of the sample depends on the accuracy of the pre-processing work, including fixation, dehydration, embedding, etc.;
3) Model matching: suitable steel knife or disposable blade; carefully consider the sample type, size and thickness when slicing, taking into account the convenience and comfort of the operator;
4) The operator’s rich experience is essential: the operator must understand the working principle of the slicer and fine-tune the machine to achieve the best results; be able to deal with common failures and eliminate some potential influencing factors;
5) Elimination of common faults: such as the angle of the slicer blade, the temperature and hardness of the embedding block, whether there are notches or impurities attached to the blade, etc.
This issue mainly talks about the knowledge of tissue sectioning. Time is limited, so I have to say goodbye to everyone. At the same time in the next issue, there will be some tips on pathology experiments. Welcome everyone to leave a message and ask questions.
You are welcome to pay more attention to Abbkine’s official account, and there will be more scientific research surprises waiting for you. At the same time, in order to provide readers with a platform for learning biological knowledge, Abbkine’s technical experts have worked tirelessly to sort out the dry goods of a variety of high-end biological experiments, which will be published in the WeChat public account in the later period. Please continue to pay attention to Abbkine for readers who love science. No public.
Abbkine focuses on the fields of proteinology and cytology, and is committed to the innovation and research and development of various antibodies, proteins, analytical reagents and kits, in order to become a key promoter in the development of life science research, drug development and other fields. We provide you with the favorite products of protein and immune research users, from basic immunological products, such as protein extraction and quantification, to internal reference label antibodies, primary and secondary antibodies for immunological experiments; the favorite products of cell research users, from Dyes and kits for detecting cell status, organelle extraction kits, cell substructure staining and tracking and cell metabolism detection products, to cytokine and protein detection kits for cell culture, just to help your research career !
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How much do you know about tissue sections? Abbkine analyzes for you one by one
Not long ago, Yacoin organized a full-blown experimental study of immunopathology, which aroused the interest of many curious babies. Everyone left messages, and the pathology experiment also included immunohistochemistry, pathology detection and other aspects, and there was still a lot of pathology knowledge that was not shown to everyone. Seeing the questions you have left, the editor will announce them one by one on the Yacoin WeChat platform in the follow-up. In this issue, the editor will start from the shallower to the deeper, starting with the knowledge of tissue slices for everyone.
When it comes to tissue sectioning, you will have many questions. Here is an analysis of your questions one by one:
What is a tissue section?
A type of qie pian slide specimen. A thin slice of animal and plant tissues for optical or electron microscope observation. Due to different requirements, a blade can be used for freehand sectioning, or the tissue block can be embedded in paraffin or collodion or frozen at low temperature and sectioned with a microtome. Cut into 5~10 micron slices for observation by optical microscope. Ultra-thin sections cut with epoxy resin or methacrylic acid embedded tissue blocks, with a thickness of 20-50 nanometers, designed for observation under an electron microscope. The sections of root tips and stems used in general teaching are generally called paraffin sections.
What is the initial shape of the tissue specimen of the tissue section?
The size of the tissue block: the ideal volume of the tissue block is 2.0cm×2.0cm×0.3cm, so that the fixative can penetrate into the tissue quickly and evenly. However, the tissue block is ideal according to the different preparation materials and purposes The volume is also different. For example, when making pathological examinations and scientific research sections, the tissue block can be thinned by 0.1~0.2cm, which can shorten the time of fixation, dehydration and transparency. If the thickness of the teaching section is 0.3~0.5cm, the same wax can be used. Block makes more teaching slices.
How to make tissue dehydration transparent?
After the specimen is fixed and washed, the tissue contains a lot of water, and the water in the tissue block must be replaced. This process is called dehydration. Whether it is sliced with paraffin or collodion, the water in the tissue must be removed. Because water-containing tissue is incompatible with embedding materials such as paraffin wax and collodion, the commonly used dehydrating agent is a series of ethanol with different concentrations. .
The dehydration steps are: 80%, 90%, 95%, 100% ethanol of various concentrations for 2 hours, this process can be completed by the automatic control of the dehydrator.
Acetone is also a dehydrating agent with strong dehydration ability, but because of its strong dehydration ability, it has a violent contraction effect on tissues, so it is generally not used when making scientific research and teaching sections. Because ethanol, acetone, etc. are not soluble in paraffin, a solvent replacement process that can dissolve in paraffin is called transparent. Commonly used transparent agents are xylene, chloroform, methyl salicylate, etc.
How to perform wax immersion, embedding, sectioning, and patching?
(1) Trimming wax block: Depending on the size of the tissue, cut off the remaining wax at about 0.1~0.2cm at the edge of the tissue, otherwise the tissue may shrink and become uneven.
(2) Prepare slicing tools: slicing knife, writing brush, ophthalmic tweezers (curved), bleaching temperature controller
(3) Install wax block: install the repaired wax block on a metal or wooden wax holder.
(4) Install the slicing knife: Install the slicing knife on the knife table of the slicing machine, and tighten the fastening screws on the knife table to prevent vibration during slicing and maintain a certain slice thickness.
(5) The thickness of the slice: the thickness regulator of the slicer is engraved with 0~50μm or 0~25μm, and the thickness can be selected arbitrarily. The thickness of the paraffin slice is generally 4~6μm.
(6) Slice
(7) Spreading: Use ophthalmic tweezers to lift the wax tape and gently spread it on the water surface at 40~45°C. Use the tension of the water and the temperature of the water to naturally flatten the slightly wrinkled wax tape.
(8) Patches and baking sheets: After the slices are fully flattened on the constant temperature water surface, the wax slices are pulled to the middle of the slide glass and the remaining water on the slide glass is poured, and placed in a 60-65 ℃ incubator Bake the slices in the oven of the temperature controller for 15 to 30 minutes to remove the paraffin that melts the tissue gap.
The picture shows a set of experimental equipment for paraffin sectioning: wax embedding machine, slicer, and spreader.
The picture shows the standard operation method of paraffin section。
How to improve the level of tissue sectioning?
1) Pathology instrument is a necessary condition: choosing the right slicer is the primary factor of the experiment;
2) The quality of the sample to be cut is a key factor: the quality of the sample depends on the accuracy of the pre-processing work, including fixation, dehydration, embedding, etc.;
3) Model matching: suitable steel knife or disposable blade; carefully consider the sample type, size and thickness when slicing, taking into account the convenience and comfort of the operator;
4) The operator’s rich experience is essential: the operator must understand the working principle of the slicer and fine-tune the machine to achieve the best results; be able to deal with common failures and eliminate some potential influencing factors;
5) Elimination of common faults: such as the angle of the slicer blade, the temperature and hardness of the embedding block, whether there are notches or impurities attached to the blade, etc.
This issue mainly talks about the knowledge of tissue sectioning. Time is limited, so I have to say goodbye to everyone. At the same time in the next issue, there will be some tips on pathology experiments. Welcome everyone to leave a message and ask questions.
You are welcome to pay more attention to Abbkine’s official account, and there will be more scientific research surprises waiting for you. At the same time, in order to provide readers with a platform for learning biological knowledge, Abbkine’s technical experts have worked tirelessly to sort out the dry goods of a variety of high-end biological experiments, which will be published in the WeChat public account in the later period. Please continue to pay attention to Abbkine for readers who love science. No public.
Abbkine focuses on the fields of proteinology and cytology, and is committed to the innovation and research and development of various antibodies, proteins, analytical reagents and kits, in order to become a key promoter in the development of life science research, drug development and other fields. We provide you with the favorite products of protein and immune research users, from basic immunological products, such as protein extraction and quantification, to internal reference label antibodies, primary and secondary antibodies for immunological experiments; the favorite products of cell research users, from Dyes and kits for detecting cell status, organelle extraction kits, cell substructure staining and tracking and cell metabolism detection products, to cytokine and protein detection kits for cell culture, just to help your research career !
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Light up fluorescence, illuminate the road to scientific research ——I wish the Abbkine IF experimental training a successful start
On November 5, 2020, under the caring leadership of superior leaders, our company successfully carried out the on-site training of “Immunofluorescence (IF)” for all Yakin employees. This training activity is an innovative activity that integrates theory and practice and conforms to the development trend of biotechnology. It has certain reference significance for the examination of biological professional knowledge in the future. At the same time, it once again embodies the exploration of the true knowledge of scientific research by the Yakein people, and responds to the resounding slogan of “lighting up the fire of fluorescence, illuminating the road to scientific research”.
In order to successfully complete this experimental operation, since its establishment, our company has invested a lot of funds to improve the consumables and equipment of scientific research laboratories. The superior leaders are also particularly concerned about the overall development of the company’s employees, and put forward many guiding opinions on the professional knowledge learning methods of our employees. This training activity is divided into three stages to start:
The first stage, the mobilization stage: the marketing department colleagues and heads of various departments will inform employees of training related matters, and complete the theoretical knowledge training of immunofluorescence (IF) experiments.
During this training process, everyone was engaged in the whole process, studied and discussed carefully, and everyone benefited a lot.
The second stage, the demonstration stage of the immunofluorescence IF experiment operation: the pathology experiment expert, Mr. Chen Haixia, personally operated and explained, and led everyone to learn all the operation procedures and precautions of the immunofluorescence experiment. Her fluent operation technique and rigorous academic spirit It is worth learning from everyone. Every link is meticulous, presenting the wonderful moments in the operation of this experiment:
Tissue dehydration and washing
Solution configuration
Incubation of antibodies
Observe slices and take pictures
Fluorescence double standard image analysis
Store the sliced product away from light
In the third stage, colleagues from various departments will take turns in the practical training stage: Yakein colleagues actively participate in the operation of the experiment. The training is carried out smoothly in a warm and harmonious atmosphere. During this training process, everyone is in actual work. The problems encountered were exchanged, and they all expressed that they had a deeper understanding of the immunofluorescence (IF) experiment process through this training class, and the benefits were quite substantial. It was a feast of learning and pointed the direction for them.
In this experiment, the marketing department and the operation department of Yakuin professionally cooperated. The whole operation video was recorded, and professional production and subtitle pronunciation were carried out to help immunofluorescence enthusiasts around the world understand the whole process of the experiment. Carry forward the love of Yakini people for biological business and the spirit of serving global customers. The video will be in the Yakein official account and Yakein official website to meet with you in the later stage, so stay tuned~~~~~
Welcome to pay more attention to “Yinfan”, and there will be great scientific research surprises waiting for everyone. At the same time, in order to provide a platform for readers to learn biological knowledge, Abbkine’s technical experts have worked tirelessly to sort out the dry goods of a variety of high-end biological experiments, and later published them in the WeChat public account. Please continue to pay attention to Abbkine for readers who love science. the public.
Abbkine focuses on the fields of proteinology and cytology, and is committed to the innovation and research and development of various antibodies, proteins, analytical reagents and kits, in order to become a key promoter in the development of life science research, drug development and other fields. We provide you with the favorite products of protein and immune research users, from basic immunological products, such as protein extraction and quantification, to internal reference label antibodies, primary antibodies and secondary antibodies for immunological experiments; the favorite products of cell research users, from Dyes and kits for detecting cell status, organelle extraction kits, cell substructure staining and tracking and cell metabolism detection products, to cytokine and protein detection kits for cell culture, just to help your research career !
0 notes
Text
Bioadvisers shared on Biotech Advisers
Light up fluorescence, illuminate the road to scientific research ——I wish the Abbkine IF experimental training a successful start
On November 5, 2020, under the caring leadership of superior leaders, our company successfully carried out the on-site training of “Immunofluorescence (IF)” for all Yakin employees. This training activity is an innovative activity that integrates theory and practice and conforms to the development trend of biotechnology. It has certain reference significance for the examination of biological professional knowledge in the future. At the same time, it once again embodies the exploration of the true knowledge of scientific research by the Yakein people, and responds to the resounding slogan of “lighting up the fire of fluorescence, illuminating the road to scientific research”.
In order to successfully complete this experimental operation, since its establishment, our company has invested a lot of funds to improve the consumables and equipment of scientific research laboratories. The superior leaders are also particularly concerned about the overall development of the company’s employees, and put forward many guiding opinions on the professional knowledge learning methods of our employees. This training activity is divided into three stages to start:
The first stage, the mobilization stage: the marketing department colleagues and heads of various departments will inform employees of training related matters, and complete the theoretical knowledge training of immunofluorescence (IF) experiments.
During this training process, everyone was engaged in the whole process, studied and discussed carefully, and everyone benefited a lot.
The second stage, the demonstration stage of the immunofluorescence IF experiment operation: the pathology experiment expert, Mr. Chen Haixia, personally operated and explained, and led everyone to learn all the operation procedures and precautions of the immunofluorescence experiment. Her fluent operation technique and rigorous academic spirit It is worth learning from everyone. Every link is meticulous, presenting the wonderful moments in the operation of this experiment:
Tissue dehydration and washing
Solution configuration
Incubation of antibodies
Observe slices and take pictures
Fluorescence double standard image analysis
Store the sliced product away from light
In the third stage, colleagues from various departments will take turns in the practical training stage: Yakein colleagues actively participate in the operation of the experiment. The training is carried out smoothly in a warm and harmonious atmosphere. During this training process, everyone is in actual work. The problems encountered were exchanged, and they all expressed that they had a deeper understanding of the immunofluorescence (IF) experiment process through this training class, and the benefits were quite substantial. It was a feast of learning and pointed the direction for them.
In this experiment, the marketing department and the operation department of Yakuin professionally cooperated. The whole operation video was recorded, and professional production and subtitle pronunciation were carried out to help immunofluorescence enthusiasts around the world understand the whole process of the experiment. Carry forward the love of Yakini people for biological business and the spirit of serving global customers. The video will be in the Yakein official account and Yakein official website to meet with you in the later stage, so stay tuned~~~~~
Welcome to pay more attention to “Yinfan”, and there will be great scientific research surprises waiting for everyone. At the same time, in order to provide a platform for readers to learn biological knowledge, Abbkine’s technical experts have worked tirelessly to sort out the dry goods of a variety of high-end biological experiments, and later published them in the WeChat public account. Please continue to pay attention to Abbkine for readers who love science. the public.
Abbkine focuses on the fields of proteinology and cytology, and is committed to the innovation and research and development of various antibodies, proteins, analytical reagents and kits, in order to become a key promoter in the development of life science research, drug development and other fields. We provide you with the favorite products of protein and immune research users, from basic immunological products, such as protein extraction and quantification, to internal reference label antibodies, primary antibodies and secondary antibodies for immunological experiments; the favorite products of cell research users, from Dyes and kits for detecting cell status, organelle extraction kits, cell substructure staining and tracking and cell metabolism detection products, to cytokine and protein detection kits for cell culture, just to help your research career !
0 notes
Text
Bioadvisers shared on Biotech Advisers
Light up fluorescence, illuminate the road to scientific research ——I wish the Abbkine IF experimental training a successful start
On November 5, 2020, under the caring leadership of superior leaders, our company successfully carried out the on-site training of “Immunofluorescence (IF)” for all Yakin employees. This training activity is an innovative activity that integrates theory and practice and conforms to the development trend of biotechnology. It has certain reference significance for the examination of biological professional knowledge in the future. At the same time, it once again embodies the exploration of the true knowledge of scientific research by the Yakein people, and responds to the resounding slogan of “lighting up the fire of fluorescence, illuminating the road to scientific research”.
In order to successfully complete this experimental operation, since its establishment, our company has invested a lot of funds to improve the consumables and equipment of scientific research laboratories. The superior leaders are also particularly concerned about the overall development of the company’s employees, and put forward many guiding opinions on the professional knowledge learning methods of our employees. This training activity is divided into three stages to start:
The first stage, the mobilization stage: the marketing department colleagues and heads of various departments will inform employees of training related matters, and complete the theoretical knowledge training of immunofluorescence (IF) experiments.
During this training process, everyone was engaged in the whole process, studied and discussed carefully, and everyone benefited a lot.
The second stage, the demonstration stage of the immunofluorescence IF experiment operation: the pathology experiment expert, Mr. Chen Haixia, personally operated and explained, and led everyone to learn all the operation procedures and precautions of the immunofluorescence experiment. Her fluent operation technique and rigorous academic spirit It is worth learning from everyone. Every link is meticulous, presenting the wonderful moments in the operation of this experiment:
Tissue dehydration and washing
Solution configuration
Incubation of antibodies
Observe slices and take pictures
Fluorescence double standard image analysis
Store the sliced product away from light
In the third stage, colleagues from various departments will take turns in the practical training stage: Yakein colleagues actively participate in the operation of the experiment. The training is carried out smoothly in a warm and harmonious atmosphere. During this training process, everyone is in actual work. The problems encountered were exchanged, and they all expressed that they had a deeper understanding of the immunofluorescence (IF) experiment process through this training class, and the benefits were quite substantial. It was a feast of learning and pointed the direction for them.
In this experiment, the marketing department and the operation department of Yakuin professionally cooperated. The whole operation video was recorded, and professional production and subtitle pronunciation were carried out to help immunofluorescence enthusiasts around the world understand the whole process of the experiment. Carry forward the love of Yakini people for biological business and the spirit of serving global customers. The video will be in the Yakein official account and Yakein official website to meet with you in the later stage, so stay tuned~~~~~
Welcome to pay more attention to “Yinfan”, and there will be great scientific research surprises waiting for everyone. At the same time, in order to provide a platform for readers to learn biological knowledge, Abbkine’s technical experts have worked tirelessly to sort out the dry goods of a variety of high-end biological experiments, and later published them in the WeChat public account. Please continue to pay attention to Abbkine for readers who love science. the public.
Abbkine focuses on the fields of proteinology and cytology, and is committed to the innovation and research and development of various antibodies, proteins, analytical reagents and kits, in order to become a key promoter in the development of life science research, drug development and other fields. We provide you with the favorite products of protein and immune research users, from basic immunological products, such as protein extraction and quantification, to internal reference label antibodies, primary antibodies and secondary antibodies for immunological experiments; the favorite products of cell research users, from Dyes and kits for detecting cell status, organelle extraction kits, cell substructure staining and tracking and cell metabolism detection products, to cytokine and protein detection kits for cell culture, just to help your research career !
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Bioadvisers shared on Biotech Advisers
Essential guidance 2 | Fishing with Purekine "fishing gear", Protein Association "wishers hooked"
In the last issue, the editor brought you “Dry Goods | Using IP “Hook” Fishing, Protein Club “Wishers Take the Bite””, and the response was very enthusiastic. In the voice of everyone, the editor is rushing through the previous experimental notes, bringing you the second bullet of protein interaction knowledge-Pull-Down. If the protein interaction is compared to an interesting “fishing” activity, then the Purekine filler we use is a tough and slender “fishing rod” and “fishing line”. The known GST-labeled recombinant protein is sharp The strong “fish hook” and “fish bait” are the perfect combination, and our target protein is the “fish” that moves freely in the water.
The concept of Pull-Down is similar to Co-IP, and its purpose is to study proteins or ligands that bind to known bait proteins. Pull-Down aims to prove the interaction between two proteins or explore unknown proteins or molecules that can bind to the target protein. Pull-Down is different from IP or Co-IP in that it is not based on antibody-antigen interaction, not an immune response. The bait protein (or ligand) is immobilized on a solid support through a non-antibody affinity system. This immobilization can be covalently coupled to the activated microbeads, or through the receptor molecule on the support The bound affinity tag binds and thus immobilizes. For example, immobilized metal chelating affinity chromatography (IMAC) resin can be used to pull-down histidine tag bait protein. The optimization of Pull-Down needs to consider the special nature of the affinity system used. Principle of Pull-Down According to the following diagram, you can understand the principle of Pull-Down:
Comparison of Pull-Down and protein purification experiments:
Use recombinant gene expression protein, and then perform downstream protein extraction and purification experiments；
Analysis of the similarities and differences between the two experimental procedures；
Experiment type Pull-Down protein purification Solid carrier Affinity chromatography resin Affinity chromatography resin filler Carrier form In bulk Column Incubation method Shaker incubation Incubate in the purification column Type of buffer Binding Buffer
Wash Buffer
Elution Buffer
Binding Buffer
Wash Buffer
Elution Buffer
Target protein form Prey protein Recombinant protein
Types of products to be tested:
Product Category Item No. His tag protein purification tool BMR20000 GST tag protein purification tool BMR20100 MBP tag protein purification tool BMR20206 Flag tag protein purification tool BMR21504 Biotin tag protein purification tool BMR20306 Protein A Antibody purification tools BMR21604 Protein GAntibody purification tools BMR20600 Protein A/G Antibody purification tools BMR20704 Protein L Antibody purification tools BMR20800
Conclusion
The Purkine series of protein purification tools provided by Abbkine are multifunctional and suitable for a variety of protein experiments. When your target protein is a recombinant protein, the purification medium can be used for the extraction of recombinant protein, which is a protein purification experiment; when your target protein is a prey protein that interacts with the recombinant protein, this series of products can be used for prey Extraction and purification.
This issue mainly talks about the knowledge of Pull-Down, time is limited, I have to say goodbye to everyone. At the same time in the next issue, we will have exciting content to meet you all. Stay tuned~~~~~
You are welcome to pay more attention to Abbkine’s official account, and there will be more scientific research surprises waiting for you. At the same time, in order to provide readers with a platform for learning biological knowledge, Abbkine’s technical experts have worked tirelessly to sort out the dry goods of a variety of high-end biological experiments, which will be published in the WeChat public account in the later period. Please continue to pay attention to Abbkine for readers who love science. No public.
Abbkine focuses on the fields of proteinology and cytology, and is committed to the innovation and research and development of various antibodies, proteins, analytical reagents and kits, in order to become a key promoter in the development of life science research, drug development and other fields. We provide you with the favorite products of protein and immune research users, from basic immunological products, such as protein extraction and quantification, to internal reference label antibodies, primary and secondary antibodies for immunological experiments; the favorite products of cell research users, from Dyes and kits for detecting cell status, organelle extraction kits, cell substructure staining and tracking and cell metabolism detection products, to cytokine and protein detection kits for cell culture, just to help your research career !
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Bioadvisers shared on Biotech Advisers
There is finally a cure for neurodegeneration!
Professor Hilmar Bading and his team’s research focused on NMDA receptors. This receptor is an ion channel protein, activated by the neurotransmitter glutamate, allowing calcium to flow into the cell. The calcium signal plays a role in the plasticity of synapses, but it also spreads to the nucleus, where it activates a protective genetic program. The NMDA receptors activated by glutamate are located at the junctions of nerve cells and play a key role in the brain, contributing to the learning and memory process and neuroprotection. But the same receptors also exist outside the synapse. These additional synaptic NMDA receptors pose a threat because their activation can cause cell death. However, under normal circumstances, an effective cellular uptake system of glutamate ensures that these receptors are not activated, and nerve cells therefore remain intact.
But when diseases occur, for example, if certain parts of the brain cannot get enough oxygen after a stroke, the interruption of blood circulation can destroy the glutamate intake system. The level of extra-synaptic glutamate increases, thereby activating the extra-synaptic NMDA receptors. The result is nerve cell damage and death, accompanied by brain function limitations. “The increase in extra-synaptic glutamate levels does not only occur during circulatory disorders in the brain,” Professor Bading explained. “Evidence shows that the toxic properties of extra synaptic NMDA receptors play an important role in many neurodegenerative diseases.” According to the scientist, this situation is in Alzheimer’s disease and amyotrophic lateral sclerosis. It is very common even after infection with viruses or parasites.
When glutamate activates NMDA receptors in neuronal junctions, they help to build a protective barrier, and outside the synapse, NMDA receptors form a “death complex” with another ion channel protein. This protein called TRPM4 has multiple functions in the body and plays a role in the cardiovascular system and immune response. According to Heidelberg’s latest findings, TRPM4 has a toxic effect on extra-synaptic NMDA receptors.
Through molecular and protein biochemical methods, scientists have determined the interface between these two interacting proteins. Armed with this knowledge, they used structure-based searches to identify substances that could disrupt this bond, thereby destroying and inactivating the “death complex.”
This class of new inhibitors-what Heidelberg researchers call “interface inhibitors” because they disrupt the bonds formed on the interface between the extra synaptic NMDA receptor and TRPM4-have proven to be extremely effective nerves Cell protection agent. “We are studying a new therapeutic drug principle. Interface inhibitors provide us with a tool that can selectively remove the toxicity of extra-synaptic NMDA receptors,” explained Professor Bading.
Professor Bading and his team have demonstrated the effectiveness of the new inhibitor in mouse models of stroke or retinal degeneration. Therefore, there are good reasons to expect this interface inhibitor-as a broad-spectrum neuroprotective agent-to provide a treatment option for neurodegenerative diseases that are currently untreatable. “However, since new substances must first successfully pass the preclinical and clinical trials, it will take several years before they may be approved as drugs for human use.”
You are welcome to pay more attention to the Abbkine, and there will be great scientific research surprises waiting for everyone. At the same time, in order to provide readers with a platform for learning biological knowledge, Abbkine’s technical experts have, after unremitting efforts, sorted out the dry goods of a variety of high-end biological experiments, and later published them in the WeChat public account. Please continue to pay attention to Abbkine for readers who love science. the public.Abbkine focuses on the fields of proteinology and cytology, and is committed to the innovation and research and development of various antibodies, proteins, analytical reagents and kits, in order to become a key promoter in the development of life science research, drug development and other fields. We provide you with the favorite products of protein and immunological research users, from basic immunology products, such as protein extraction and quantification, to internal reference label antibodies, primary antibodies and secondary antibodies for immunological experiments; cell research users’ favorite products, from Dyes and kits for detecting cell status, organelle extraction kits, cell substructure staining and tracking and cell metabolism detection products, to cytokine and protein detection kits for cell culture, are only for your research career !
0 notes
Text
Bioadvisers shared on Biotech Advisers
There is finally a cure for neurodegeneration!
Professor Hilmar Bading and his team’s research focused on NMDA receptors. This receptor is an ion channel protein, activated by the neurotransmitter glutamate, allowing calcium to flow into the cell. The calcium signal plays a role in the plasticity of synapses, but it also spreads to the nucleus, where it activates a protective genetic program. The NMDA receptors activated by glutamate are located at the junctions of nerve cells and play a key role in the brain, contributing to the learning and memory process and neuroprotection. But the same receptors also exist outside the synapse. These additional synaptic NMDA receptors pose a threat because their activation can cause cell death. However, under normal circumstances, an effective cellular uptake system of glutamate ensures that these receptors are not activated, and nerve cells therefore remain intact.
But when diseases occur, for example, if certain parts of the brain cannot get enough oxygen after a stroke, the interruption of blood circulation can destroy the glutamate intake system. The level of extra-synaptic glutamate increases, thereby activating the extra-synaptic NMDA receptors. The result is nerve cell damage and death, accompanied by brain function limitations. “The increase in extra-synaptic glutamate levels does not only occur during circulatory disorders in the brain,” Professor Bading explained. “Evidence shows that the toxic properties of extra synaptic NMDA receptors play an important role in many neurodegenerative diseases.” According to the scientist, this situation is in Alzheimer’s disease and amyotrophic lateral sclerosis. It is very common even after infection with viruses or parasites.
When glutamate activates NMDA receptors in neuronal junctions, they help to build a protective barrier, and outside the synapse, NMDA receptors form a “death complex” with another ion channel protein. This protein called TRPM4 has multiple functions in the body and plays a role in the cardiovascular system and immune response. According to Heidelberg’s latest findings, TRPM4 has a toxic effect on extra-synaptic NMDA receptors.
Through molecular and protein biochemical methods, scientists have determined the interface between these two interacting proteins. Armed with this knowledge, they used structure-based searches to identify substances that could disrupt this bond, thereby destroying and inactivating the “death complex.”
This class of new inhibitors-what Heidelberg researchers call “interface inhibitors” because they disrupt the bonds formed on the interface between the extra synaptic NMDA receptor and TRPM4-have proven to be extremely effective nerves Cell protection agent. “We are studying a new therapeutic drug principle. Interface inhibitors provide us with a tool that can selectively remove the toxicity of extra-synaptic NMDA receptors,” explained Professor Bading.
Professor Bading and his team have demonstrated the effectiveness of the new inhibitor in mouse models of stroke or retinal degeneration. Therefore, there are good reasons to expect this interface inhibitor-as a broad-spectrum neuroprotective agent-to provide a treatment option for neurodegenerative diseases that are currently untreatable. “However, since new substances must first successfully pass the preclinical and clinical trials, it will take several years before they may be approved as drugs for human use.”
You are welcome to pay more attention to the Abbkine, and there will be great scientific research surprises waiting for everyone. At the same time, in order to provide readers with a platform for learning biological knowledge, Abbkine’s technical experts have, after unremitting efforts, sorted out the dry goods of a variety of high-end biological experiments, and later published them in the WeChat public account. Please continue to pay attention to Abbkine for readers who love science. the public.Abbkine focuses on the fields of proteinology and cytology, and is committed to the innovation and research and development of various antibodies, proteins, analytical reagents and kits, in order to become a key promoter in the development of life science research, drug development and other fields. We provide you with the favorite products of protein and immunological research users, from basic immunology products, such as protein extraction and quantification, to internal reference label antibodies, primary antibodies and secondary antibodies for immunological experiments; cell research users’ favorite products, from Dyes and kits for detecting cell status, organelle extraction kits, cell substructure staining and tracking and cell metabolism detection products, to cytokine and protein detection kits for cell culture, are only for your research career !
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Bioadvisers shared on Biotech Advisers
Using immunoprecipitation "fish hooks" for fishing, the protein will always "hooked with willing"
The thoughtful Thursday is here again, and the editor who has long been unable to bear it is finally released to meet our favorite “Yin Fans”.
This time, the editor brings you a wave of colorful biological science mysteries-protein interaction. If protein interaction is compared to an interesting “fishing” activity, then the tag antibody coupling carrier we use is a sharp and strong “fish hook”, and the tag recombinant protein (bait protein) is sweet and delicious. “Bait”, and our target protein is the “fish” that moves freely in the water.
If you want to do your job well, you must first sharpen your tools. Maybe you are still worrying about the purification and enrichment of the target protein without a good antibody coupled to Beads. Maybe the protein content you collected at this time is low, maybe you are enriching it at this time. The protein impurity content is high, maybe the protein vitality that you have harvested through the hard work at this time has been reduced, maybe the Beads operation steps you are using now are long and take a lot of time, maybe, maybe…
Next, the editor will give you a feast of protein experiments: reveal the whole process of protein interaction, what kind of magic operation is IP and Co-IP?
Immunoprecipitation (IP)
“Immunoprecipitation” generally refers to a small-scale protein affinity purification experiment using a binding protein immobilized on a solid support. More precisely, IP is an experiment in which a specific antibody immobilized on a microbead support (usually agarose resin) is used to purify a single antigen from a complex mixture. The assembly of the immobilized protein complex can be performed step by step or completed in one step (Figure 1). Common loading sequence: incubate the antibody and sample (such as cell lysate) together, and then add affinity beads to capture the antibody-antigen complex. It is also possible to incubate the antibody and microbeads (directly or indirectly bind to the antibody through the antibody binding protein, such as protein A, G, or AG), and then add the sample containing the antigen. After the antigen, antibody and solid support are bound, the beads are washed thoroughly, and then an appropriate elution buffer is used to elute the antigen from the support.
Co-immunoprecipitation(Co-IP)
Co-immunoprecipitation analysis (Co-IP) is very similar to IP. The basic technique is to use immobilized antibodies specific to the target antigen; however, the goal of IP is to purify a single antigen, while Co-IP is designed to separate the antigen and the antigen. Bound protein or ligand. In the Co-IP experiment, the known antigen is called the bait protein, and the protein bound to it is called the target protein. The target protein may be some complex chaperone proteins, signal molecules, structural proteins, cofactors, etc. The intensity of the interaction between proteins may range between highly transient and very stable. The basic Co-IP experiment scheme is the same as that of IP. In fact, any IP system can be used for Co-IP. However, there are many other factors that need to be considered, for example, the optimization of binding and washing conditions. When optimizing, the strength of the bait protein-target protein interaction and the affinity of the antibody-bait protein need to be considered.
Disadvantages of Co-IP experiment:
Low-affinity and instant protein-protein interactions may not be detected; the binding of the two proteins may not be a direct binding, but a third party may act as a bridge in the middle; the target protein must be predicted before the experiment to choose the final Antibodies tested, so if the prediction is incorrect, the experiment will have no results.
Advantages of Co-IP experiment:
The interacting proteins are all post-translationally modified and are in a natural state; protein interactions are carried out in a natural mixed protein state, which can avoid human influence; the interacting protein complex in the natural state can be separated.
Technical upgrade of IP and Co-IP
Basic steps of traditional IP and Co-IP:
Add the appropriate antibody.
The antibody binds to the target protein.
Add Protein A/G to make the antibody-protein complex insoluble.
Centrifuge the solution to separate the antibody-protein complex.
Remove the supernatant and wash.
Limitations of traditional ProteinA/G in IP applications:
ü1) Time-consuming
ü2) When the antibody species, subtype and ProteinA/G do not match, the ability of ProteinA/G to bind to the antibody is weak
ü3) Binding with other immunoglobulins in the sample, resulting in non-specific binding, resulting in high background
Basic steps of IP and Co-IP after upgrade:
Add the appropriate antibody conjugate.
The antibody binds to the target protein.
Centrifuge the solution to separate the antibody-protein complex.
Remove the supernatant and wash.
After using our upgraded agarose/magnetic bead coupling tag antibody, we can be surprised to find that the experimental process eliminates a time-consuming and complicated experimental operation of antibody and carrier binding, and the perfect combination of antibody and Beads , Improve the accuracy of the target protein being “pulled”, the purity and yield of the target protein obtained will be significantly improved, and the operation is simple, saving time and effort!
Having said that, you must be impatient for our Abbkine’s “protein weapon”. The editor is now very proud to recommend our agarose/magnetic bead conjugated antibody to everyone:
Cat.NO Product name Application A02010AGB Anti-DDDDK Tag Mouse Monoclonal Antibody, Agarose) IP/Co-IP A02010MGB Anti-DDDDK Tag Mouse Monoclonal Antibody, Magnetic Beads) IP/Co-IP A02040AGB Anti-HA Tag Mouse Monoclonal Antibody, Agarose IP/Co-IP A02040MGB Anti-HA Tag Mouse Monoclonal Antibody, Magnetic Beads IP/Co-IP A02050AGB Anti-His Tag Mouse Monoclonal Antibody, Agarose IP/Co-IP A02050MGB Anti-His Tag Mouse Monoclonal Antibody, Magnetic Beads IP/Co-IP A02060AGB Anti-Myc Tag Mouse Monoclonal Antibody, Agarose IP/Co-IP A02060MGB Anti-Myc Tag Mouse Monoclonal Antibody, Magnetic Beads IP/Co-IP A02170AGB Anti-V5 Tag Mouse Monoclonal Antibody, Agarose IP/Co-IP A02170MGB Anti-V5 Tag Mouse Monoclonal Antibody, Magnetic Beads IP/Co-IP
This issue is mainly about the knowledge of immunoprecipitation, time is limited, I have to say goodbye to everyone again. At the same time in the next issue, there will be Pull down operation tips with protein interaction to meet you. Stay continued~~~~~
You are welcome to pay more attention to the Abbkine, and there will be great scientific research surprises waiting for everyone. At the same time, in order to provide readers with a platform for learning biological knowledge, Abbkine’s technical experts have, after unremitting efforts, sorted out the dry goods of a variety of high-end biological experiments, and later published them in the WeChat public account. Please continue to pay attention to Abbkine for readers who love science. the public.
Abbkine focuses on the fields of proteinology and cytology, and is committed to the innovation and research and development of various antibodies, proteins, analytical reagents and kits, in order to become a key promoter in the development of life science research, drug development and other fields. We provide you with the favorite products of protein and immunological research users, from basic immunology products, such as protein extraction and quantification, to internal reference label antibodies, primary antibodies and secondary antibodies for immunological experiments; cell research users’ favorite products, from Dyes and kits for detecting cell status, organelle extraction kits, cell substructure staining and tracking and cell metabolism detection products, to cytokine and protein detection kits for cell culture, are only for your research career !
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A Common test of Hydrogen Peroxide
Hydrogen Peroxide is a peroxide and oxidizing agent with disinfectant, antiviral and anti-bacterial activities. Upon rinsing and gargling or topical application, hydrogen peroxide exerts its oxidizing activity and produces free radicals which leads to oxidative damage to proteins and membrane lipids. This may inactivate and destroy pathogens and may prevent spreading of infection.
Hydrogen peroxide is unstable and slowly decomposes in the presence of light. Because of its instability, hydrogen peroxide is typically stored with a stabilizer in a weakly acidic solution in a dark coloured bottle. Hydrogen peroxide is found in biological systems including the human body. Enzymes that use or decompose hydrogen peroxide are classified as peroxidases.
Fig 1: The Role of Hydrogen Peroxide in Redox-Dependent Signaling
(Picture Source: https://www.mdpi.com/2073-4409/7/10/156/htm)
Through year of efforts, Abbkine research scientists have developed a series of assay kits for cell metabolism and that constitutes Abbkine CheKine™group. For detailed index, you are welcome to view at https://www.abbkine.com/?s=CheKine&post_type=product&submit= . In which we can see the detection kits of NAD/NADH, NADP/NADPH, superoxide dismutases (SOD), peroxidase (HRP) and glucose, but today we are going to introduce the test of H2O2 for you.
To more specific, the benefits of Abbkine Hydrogen Peroxide (H2O2) Assay Kit can be concluded as below:
Determination of Hydrogen Peroxide in serum, plasma, cell/tissue samples, urine and other biological fluids.
Detect biological samples at a level of 1-20 µM.
Simple procedures, fase and convenient, and more stable.
You can view the kit protocol at https://www.abbkine.com/file/booklet/KTB1041-B.pdf .
Fig 2: Standard Curve of Hydrogen Peroxide Assay.
Kit Components:
Reaction Buffer
H2O2 Standard (1M)
Assay Buffer (10X)
Abbkine is aimed at to provide high quality and cost effective reagents to global scientists, this Hydrogen Peroxide (H2O2) Assay Kit has been optimized to use and can deduct your experiment cost. For detailed product information, you can refer in below sheet.
CAT# Product Name Size and Price KTB1041 CheKine™ Hydrogen Peroxide (H2O2) Assay Kit
$40/48T
$60/96T
If you are interested in Abbkine CheKine™Hydrogen Peroxide (H2O2) Assay Kit, feel free to contact [email protected], our service representative will get back to you shortly!
About Abbkine Scientific Co., Ltd.
Abbkine offers services for global scientists in the fields of proteomics and cytology, and is committed to the innovation and development of various scientific reagents related to proteomics and cytology, to promote the development of fields including life science research and drug discovery. Proteomics products cover the preparation of samples (protein extraction, purification, coupling), protein quantification, antibodies and kits for protein detection. Cytology products involve cytokines (cell culture), cell status detection, cell staining, organelle extraction, cell metabolism and cytopathology reagents (kits). Abbkine relies on the product portfolio and unique marketing support as the main market strategy and product innovation mode, to help your research career!
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