seen from Türkiye
seen from Mexico
seen from Italy
seen from Thailand
seen from United States
seen from United States
seen from Brazil
seen from Belgium
seen from Singapore
seen from Indonesia
seen from Australia
seen from Taiwan
seen from Malaysia
seen from Taiwan
seen from United Kingdom
seen from China
seen from United States
seen from China
seen from Norway
seen from China
My PI just handed me a protocol from his post doc written on a paper towel. Yes, I did put it in a sheet protector!
BioAdvisers said on Biotech Advisers
WB Essential guidance| After returning to school you must know the product portfolio of WB protein analysis
Following the previous issue of “WB Essential guidance | WB Sample Pre-Processing Product Portfolio Scheme that You Must Know After Back to School”, the little friends have a deep understanding of the sample pre-processing after learning. The editor here also received the ardent new and old customers of Abbkine eager for Abbkine products. Therefore, the second part of WB dry goods series: After returning to school, you must know the product portfolio plan after WB transfer film, which came into being, and continue to answer questions for everyone.
In the last issue, we talked about protein quantification products. Everyone may be curious, how should protein quantification products be selected? Next announced for everyone:
The most popular methods for protein quantification are BCA and Bradford:
BCA method: tolerant to detergents: SDS, TritonX-100, Tween-20, etc., but not to reducing agents, if the sample contains detergents without reducing agents, this method can be used (recommended KTD3001)
Bradford method: can tolerate the chemicals in most samples, not high concentration detergent, if the sample contains high concentration detergent, this method can be used (recommended KTD3002)
After protein quantification, doing protein analysis is naturally the most concern of WB experimenters.
The first step is to load the sample. In the sample loading process, in addition to the sample order that you cannot forget, the most important thing is the protein marker. In order to clearly indicate the molecular weight of the protein, some senior WB operators will choose the point 2-3 pre-stained protein wells to avoid experimental uncertainty and improve the clarity of instructions. Here we recommend two color pre-stained protein markers:
From the experimental results, the BMM3001 and BMM3002 markers have bright colors, and the sharpness and contrast are very high. The reference value of the indication result is very high.
After a long and lengthy step of running rubber and transferring membrane, it finally entered the stage of WB immunization. At this time, how to correctly select the internal reference antibody will become an important step in the WB experiment. The following dry goods are here again-the three magic weapons for the correct internal reference selection:
Species source: mammalian tissue or cell samples, usually choose β-actin, β-tubulin, GAPDH, etc., plant experiment samples use plant actin, Rubisco, etc.
Protein molecular weight: The molecular weight of the target protein should be considered. Generally, the molecular weight difference between the target protein and the internal reference protein should be more than 5KD; for high molecular weight proteins over 100 kDa, Vinculin focal adhesion protein is recommended as the internal reference antibody.
Protein expression site: According to the different protein expression positions, more suitable antibodies can be selected. Commonly used extensive β-actin, β-tubulin, GAPDH; detection of nuclear Lamin A, Histone H3, PCNA, detection of cell membrane Na, K ATPase, etc.
In addition to free antibody, Abbkine’s internal reference antibody also has a straight-labeled antibody conjugated with a label. If you think it is not enough, you can also contact our Abbine technical experts ([email protected]) for in-depth communication. We will help you find the most suitable internal control antibody for your experiment.
In addition to internal reference antibodies, the selection of target protein antibodies is the finishing touch of our WB experiment. At this point, you may consider: how to select antibodies with strong specificity, high sensitivity, and more literature support? The first choice is Abbkine’s first antibody. After hearing the name, we know that our antibody has been carefully tested by Abbkine scientists on the functional properties of the antibody. So the customers who used it are full of praise. Do not believe? You can also try.
Of course, Abbkine’s common monoclonal antibodies and polyclonal antibodies are also effective and cost-effective. Used children’s shoes are said to be good. If the customer’s sample is a recombinant protein with a protein tag, we can also provide the corresponding labeled antibody and its direct-labeled antibody to meet the needs of customers with a variety of experiments.
Product NO Product Name A02010 Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10) A02020 Anti-GFP Tag Mouse Monoclonal Antibody (3D3) A02030 Anti-GST Tag Mouse Monoclonal Antibody (2A8) A02040 Anti-HA Tag Mouse Monoclonal Antibody (4F6) A02050 Anti-His Tag Mouse Monoclonal Antibody (5C3) A02060 Anti-Myc Tag Mouse Monoclonal Antibody (2D5)
Of course, many customers are interested in the coupling kits independently developed by Abbkine. Our company also has carefully created the LIKINE series of kits for you. The self-conjugated antibodies will also be very fulfilling to use.AbFluor™ dyes are a series of highly water-soluble fluorescent dyes spanning full UV-visible and near-infrared spectrum for labeling biomolecules, especially proteins and nucleic acids. AbFluor™ dyes have much better labeling performance than other fluorescent labeling dyes such as FITC, TRITC, Cy® dyes, Alexa Fluor® and Dylight® labeling dyes.
At the end of the experiment, of course, the use of color developing solution, Abckine’s ECL color developing solution can reach the pico level and fly level. This sensitivity is very impressive. SuperLumia high-sensitivity and ultra-sensitive ECL products have extremely high sensitivity, and require less samples and antibodies; the luminous signal is strong and lasts for a long time.
Product No Product Name K22020 SuperLumia ECL HRP Substrate Kit K22030 SuperLumia ECL Plus HRP Substrate Kit
In order to provide readers with a platform for learning biological knowledge, Abbkine’s technical experts have worked hard to sort out the dry goods of various high-end biological experiments. Later, they will be published in the WeChat public account, and readers who love science will continue to pay attention to the Abbkine public account. .
Abbkine specializes in the fields of proteinology and cytology, and is committed to innovating and developing various antibodies, proteins, analytical reagents and kits, with a view to becoming a key promoter in the fields of life science research and development, drug research and development. We provide you with favorite products of protein and immunological research users, from basic immunological products, such as protein extraction and quantification, to internal reference labeling antibodies, primary antibodies and secondary antibodies of immunological experiments, etc.; favorite products of cell research users, from Dyes and kits for detecting the state of cells, kits for extracting organelles, staining and tracking of cell substructures and cell metabolism detection products, to cytokine and protein detection kits for cell culture, only to help your research career !
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.
Bioadvisers shared on Biotech Advisers
WB Essential guidance| After returning to school you must know the product portfolio of WB protein analysis
Following the previous issue of “WB Essential guidance | WB Sample Pre-Processing Product Portfolio Scheme that You Must Know After Back to School”, the little friends have a deep understanding of the sample pre-processing after learning. The editor here also received the ardent new and old customers of Abbkine eager for Abbkine products. Therefore, the second part of WB dry goods series: After returning to school, you must know the product portfolio plan after WB transfer film, which came into being, and continue to answer questions for everyone.
In the last issue, we talked about protein quantification products. Everyone may be curious, how should protein quantification products be selected? Next announced for everyone:
The most popular methods for protein quantification are BCA and Bradford:
BCA method: tolerant to detergents: SDS, TritonX-100, Tween-20, etc., but not to reducing agents, if the sample contains detergents without reducing agents, this method can be used (recommended KTD3001)
Bradford method: can tolerate the chemicals in most samples, not high concentration detergent, if the sample contains high concentration detergent, this method can be used (recommended KTD3002)
After protein quantification, doing protein analysis is naturally the most concern of WB experimenters.
The first step is to load the sample. In the sample loading process, in addition to the sample order that you cannot forget, the most important thing is the protein marker. In order to clearly indicate the molecular weight of the protein, some senior WB operators will choose the point 2-3 pre-stained protein wells to avoid experimental uncertainty and improve the clarity of instructions. Here we recommend two color pre-stained protein markers:
From the experimental results, the BMM3001 and BMM3002 markers have bright colors, and the sharpness and contrast are very high. The reference value of the indication result is very high.
After a long and lengthy step of running rubber and transferring membrane, it finally entered the stage of WB immunization. At this time, how to correctly select the internal reference antibody will become an important step in the WB experiment. The following dry goods are here again-the three magic weapons for the correct internal reference selection:
Species source: mammalian tissue or cell samples, usually choose β-actin, β-tubulin, GAPDH, etc., plant experiment samples use plant actin, Rubisco, etc.
Protein molecular weight: The molecular weight of the target protein should be considered. Generally, the molecular weight difference between the target protein and the internal reference protein should be more than 5KD; for high molecular weight proteins over 100 kDa, Vinculin focal adhesion protein is recommended as the internal reference antibody.
Protein expression site: According to the different protein expression positions, more suitable antibodies can be selected. Commonly used extensive β-actin, β-tubulin, GAPDH; detection of nuclear Lamin A, Histone H3, PCNA, detection of cell membrane Na, K ATPase, etc.
In addition to free antibody, Abbkine’s internal reference antibody also has a straight-labeled antibody conjugated with a label. If you think it is not enough, you can also contact our Abbine technical experts ([email protected]) for in-depth communication. We will help you find the most suitable internal control antibody for your experiment.
In addition to internal reference antibodies, the selection of target protein antibodies is the finishing touch of our WB experiment. At this point, you may consider: how to select antibodies with strong specificity, high sensitivity, and more literature support? The first choice is Abbkine’s first antibody. After hearing the name, we know that our antibody has been carefully tested by Abbkine scientists on the functional properties of the antibody. So the customers who used it are full of praise. Do not believe? You can also try.
Of course, Abbkine’s common monoclonal antibodies and polyclonal antibodies are also effective and cost-effective. Used children’s shoes are said to be good. If the customer’s sample is a recombinant protein with a protein tag, we can also provide the corresponding labeled antibody and its direct-labeled antibody to meet the needs of customers with a variety of experiments.
Product NO Product Name A02010 Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10) A02020 Anti-GFP Tag Mouse Monoclonal Antibody (3D3) A02030 Anti-GST Tag Mouse Monoclonal Antibody (2A8) A02040 Anti-HA Tag Mouse Monoclonal Antibody (4F6) A02050 Anti-His Tag Mouse Monoclonal Antibody (5C3) A02060 Anti-Myc Tag Mouse Monoclonal Antibody (2D5)
Of course, many customers are interested in the coupling kits independently developed by Abbkine. Our company also has carefully created the LIKINE series of kits for you. The self-conjugated antibodies will also be very fulfilling to use.AbFluor™ dyes are a series of highly water-soluble fluorescent dyes spanning full UV-visible and near-infrared spectrum for labeling biomolecules, especially proteins and nucleic acids. AbFluor™ dyes have much better labeling performance than other fluorescent labeling dyes such as FITC, TRITC, Cy® dyes, Alexa Fluor® and Dylight® labeling dyes.
At the end of the experiment, of course, the use of color developing solution, Abckine’s ECL color developing solution can reach the pico level and fly level. This sensitivity is very impressive. SuperLumia high-sensitivity and ultra-sensitive ECL products have extremely high sensitivity, and require less samples and antibodies; the luminous signal is strong and lasts for a long time.
Product No Product Name K22020 SuperLumia ECL HRP Substrate Kit K22030 SuperLumia ECL Plus HRP Substrate Kit
In order to provide readers with a platform for learning biological knowledge, Abbkine’s technical experts have worked hard to sort out the dry goods of various high-end biological experiments. Later, they will be published in the WeChat public account, and readers who love science will continue to pay attention to the Abbkine public account. .
Abbkine specializes in the fields of proteinology and cytology, and is committed to innovating and developing various antibodies, proteins, analytical reagents and kits, with a view to becoming a key promoter in the fields of life science research and development, drug research and development. We provide you with favorite products of protein and immunological research users, from basic immunological products, such as protein extraction and quantification, to internal reference labeling antibodies, primary antibodies and secondary antibodies of immunological experiments, etc.; favorite products of cell research users, from Dyes and kits for detecting the state of cells, kits for extracting organelles, staining and tracking of cell substructures and cell metabolism detection products, to cytokine and protein detection kits for cell culture, only to help your research career !
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.
Bioadvisers shared on Biotech Advisers
WB Essential guidance| After returning to school you must know the product portfolio of WB protein analysis
Following the previous issue of “WB Essential guidance | WB Sample Pre-Processing Product Portfolio Scheme that You Must Know After Back to School”, the little friends have a deep understanding of the sample pre-processing after learning. The editor here also received the ardent new and old customers of Abbkine eager for Abbkine products. Therefore, the second part of WB dry goods series: After returning to school, you must know the product portfolio plan after WB transfer film, which came into being, and continue to answer questions for everyone.
In the last issue, we talked about protein quantification products. Everyone may be curious, how should protein quantification products be selected? Next announced for everyone:
The most popular methods for protein quantification are BCA and Bradford:
BCA method: tolerant to detergents: SDS, TritonX-100, Tween-20, etc., but not to reducing agents, if the sample contains detergents without reducing agents, this method can be used (recommended KTD3001)
Bradford method: can tolerate the chemicals in most samples, not high concentration detergent, if the sample contains high concentration detergent, this method can be used (recommended KTD3002)
After protein quantification, doing protein analysis is naturally the most concern of WB experimenters.
The first step is to load the sample. In the sample loading process, in addition to the sample order that you cannot forget, the most important thing is the protein marker. In order to clearly indicate the molecular weight of the protein, some senior WB operators will choose the point 2-3 pre-stained protein wells to avoid experimental uncertainty and improve the clarity of instructions. Here we recommend two color pre-stained protein markers:
From the experimental results, the BMM3001 and BMM3002 markers have bright colors, and the sharpness and contrast are very high. The reference value of the indication result is very high.
After a long and lengthy step of running rubber and transferring membrane, it finally entered the stage of WB immunization. At this time, how to correctly select the internal reference antibody will become an important step in the WB experiment. The following dry goods are here again-the three magic weapons for the correct internal reference selection:
Species source: mammalian tissue or cell samples, usually choose β-actin, β-tubulin, GAPDH, etc., plant experiment samples use plant actin, Rubisco, etc.
Protein molecular weight: The molecular weight of the target protein should be considered. Generally, the molecular weight difference between the target protein and the internal reference protein should be more than 5KD; for high molecular weight proteins over 100 kDa, Vinculin focal adhesion protein is recommended as the internal reference antibody.
Protein expression site: According to the different protein expression positions, more suitable antibodies can be selected. Commonly used extensive β-actin, β-tubulin, GAPDH; detection of nuclear Lamin A, Histone H3, PCNA, detection of cell membrane Na, K ATPase, etc.
In addition to free antibody, Abbkine’s internal reference antibody also has a straight-labeled antibody conjugated with a label. If you think it is not enough, you can also contact our Abbine technical experts ([email protected]) for in-depth communication. We will help you find the most suitable internal control antibody for your experiment.
In addition to internal reference antibodies, the selection of target protein antibodies is the finishing touch of our WB experiment. At this point, you may consider: how to select antibodies with strong specificity, high sensitivity, and more literature support? The first choice is Abbkine’s first antibody. After hearing the name, we know that our antibody has been carefully tested by Abbkine scientists on the functional properties of the antibody. So the customers who used it are full of praise. Do not believe? You can also try.
Of course, Abbkine’s common monoclonal antibodies and polyclonal antibodies are also effective and cost-effective. Used children’s shoes are said to be good. If the customer’s sample is a recombinant protein with a protein tag, we can also provide the corresponding labeled antibody and its direct-labeled antibody to meet the needs of customers with a variety of experiments.
Product NO Product Name A02010 Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10) A02020 Anti-GFP Tag Mouse Monoclonal Antibody (3D3) A02030 Anti-GST Tag Mouse Monoclonal Antibody (2A8) A02040 Anti-HA Tag Mouse Monoclonal Antibody (4F6) A02050 Anti-His Tag Mouse Monoclonal Antibody (5C3) A02060 Anti-Myc Tag Mouse Monoclonal Antibody (2D5)
Of course, many customers are interested in the coupling kits independently developed by Abbkine. Our company also has carefully created the LIKINE series of kits for you. The self-conjugated antibodies will also be very fulfilling to use.AbFluor™ dyes are a series of highly water-soluble fluorescent dyes spanning full UV-visible and near-infrared spectrum for labeling biomolecules, especially proteins and nucleic acids. AbFluor™ dyes have much better labeling performance than other fluorescent labeling dyes such as FITC, TRITC, Cy® dyes, Alexa Fluor® and Dylight® labeling dyes.
At the end of the experiment, of course, the use of color developing solution, Abckine’s ECL color developing solution can reach the pico level and fly level. This sensitivity is very impressive. SuperLumia high-sensitivity and ultra-sensitive ECL products have extremely high sensitivity, and require less samples and antibodies; the luminous signal is strong and lasts for a long time.
Product No Product Name K22020 SuperLumia ECL HRP Substrate Kit K22030 SuperLumia ECL Plus HRP Substrate Kit
In order to provide readers with a platform for learning biological knowledge, Abbkine’s technical experts have worked hard to sort out the dry goods of various high-end biological experiments. Later, they will be published in the WeChat public account, and readers who love science will continue to pay attention to the Abbkine public account. .
Abbkine specializes in the fields of proteinology and cytology, and is committed to innovating and developing various antibodies, proteins, analytical reagents and kits, with a view to becoming a key promoter in the fields of life science research and development, drug research and development. We provide you with favorite products of protein and immunological research users, from basic immunological products, such as protein extraction and quantification, to internal reference labeling antibodies, primary antibodies and secondary antibodies of immunological experiments, etc.; favorite products of cell research users, from Dyes and kits for detecting the state of cells, kits for extracting organelles, staining and tracking of cell substructures and cell metabolism detection products, to cytokine and protein detection kits for cell culture, only to help your research career !
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.
Bioadvisers shared on Biotech Advisers
WB Essential guidance| After returning to school you must know the product portfolio of WB protein analysis
Following the previous issue of “WB Essential guidance | WB Sample Pre-Processing Product Portfolio Scheme that You Must Know After Back to School”, the little friends have a deep understanding of the sample pre-processing after learning. The editor here also received the ardent new and old customers of Abbkine eager for Abbkine products. Therefore, the second part of WB dry goods series: After returning to school, you must know the product portfolio plan after WB transfer film, which came into being, and continue to answer questions for everyone.
In the last issue, we talked about protein quantification products. Everyone may be curious, how should protein quantification products be selected? Next announced for everyone:
The most popular methods for protein quantification are BCA and Bradford:
BCA method: tolerant to detergents: SDS, TritonX-100, Tween-20, etc., but not to reducing agents, if the sample contains detergents without reducing agents, this method can be used (recommended KTD3001)
Bradford method: can tolerate the chemicals in most samples, not high concentration detergent, if the sample contains high concentration detergent, this method can be used (recommended KTD3002)
After protein quantification, doing protein analysis is naturally the most concern of WB experimenters.
The first step is to load the sample. In the sample loading process, in addition to the sample order that you cannot forget, the most important thing is the protein marker. In order to clearly indicate the molecular weight of the protein, some senior WB operators will choose the point 2-3 pre-stained protein wells to avoid experimental uncertainty and improve the clarity of instructions. Here we recommend two color pre-stained protein markers:
From the experimental results, the BMM3001 and BMM3002 markers have bright colors, and the sharpness and contrast are very high. The reference value of the indication result is very high.
After a long and lengthy step of running rubber and transferring membrane, it finally entered the stage of WB immunization. At this time, how to correctly select the internal reference antibody will become an important step in the WB experiment. The following dry goods are here again-the three magic weapons for the correct internal reference selection:
Species source: mammalian tissue or cell samples, usually choose β-actin, β-tubulin, GAPDH, etc., plant experiment samples use plant actin, Rubisco, etc.
Protein molecular weight: The molecular weight of the target protein should be considered. Generally, the molecular weight difference between the target protein and the internal reference protein should be more than 5KD; for high molecular weight proteins over 100 kDa, Vinculin focal adhesion protein is recommended as the internal reference antibody.
Protein expression site: According to the different protein expression positions, more suitable antibodies can be selected. Commonly used extensive β-actin, β-tubulin, GAPDH; detection of nuclear Lamin A, Histone H3, PCNA, detection of cell membrane Na, K ATPase, etc.
In addition to free antibody, Abbkine’s internal reference antibody also has a straight-labeled antibody conjugated with a label. If you think it is not enough, you can also contact our Abbine technical experts ([email protected]) for in-depth communication. We will help you find the most suitable internal control antibody for your experiment.
In addition to internal reference antibodies, the selection of target protein antibodies is the finishing touch of our WB experiment. At this point, you may consider: how to select antibodies with strong specificity, high sensitivity, and more literature support? The first choice is Abbkine’s first antibody. After hearing the name, we know that our antibody has been carefully tested by Abbkine scientists on the functional properties of the antibody. So the customers who used it are full of praise. Do not believe? You can also try.
Of course, Abbkine’s common monoclonal antibodies and polyclonal antibodies are also effective and cost-effective. Used children’s shoes are said to be good. If the customer’s sample is a recombinant protein with a protein tag, we can also provide the corresponding labeled antibody and its direct-labeled antibody to meet the needs of customers with a variety of experiments.
Product NO Product Name A02010 Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10) A02020 Anti-GFP Tag Mouse Monoclonal Antibody (3D3) A02030 Anti-GST Tag Mouse Monoclonal Antibody (2A8) A02040 Anti-HA Tag Mouse Monoclonal Antibody (4F6) A02050 Anti-His Tag Mouse Monoclonal Antibody (5C3) A02060 Anti-Myc Tag Mouse Monoclonal Antibody (2D5)
Of course, many customers are interested in the coupling kits independently developed by Abbkine. Our company also has carefully created the LIKINE series of kits for you. The self-conjugated antibodies will also be very fulfilling to use.AbFluor™ dyes are a series of highly water-soluble fluorescent dyes spanning full UV-visible and near-infrared spectrum for labeling biomolecules, especially proteins and nucleic acids. AbFluor™ dyes have much better labeling performance than other fluorescent labeling dyes such as FITC, TRITC, Cy® dyes, Alexa Fluor® and Dylight® labeling dyes.
At the end of the experiment, of course, the use of color developing solution, Abckine’s ECL color developing solution can reach the pico level and fly level. This sensitivity is very impressive. SuperLumia high-sensitivity and ultra-sensitive ECL products have extremely high sensitivity, and require less samples and antibodies; the luminous signal is strong and lasts for a long time.
Product No Product Name K22020 SuperLumia ECL HRP Substrate Kit K22030 SuperLumia ECL Plus HRP Substrate Kit
In order to provide readers with a platform for learning biological knowledge, Abbkine’s technical experts have worked hard to sort out the dry goods of various high-end biological experiments. Later, they will be published in the WeChat public account, and readers who love science will continue to pay attention to the Abbkine public account. .
Abbkine specializes in the fields of proteinology and cytology, and is committed to innovating and developing various antibodies, proteins, analytical reagents and kits, with a view to becoming a key promoter in the fields of life science research and development, drug research and development. We provide you with favorite products of protein and immunological research users, from basic immunological products, such as protein extraction and quantification, to internal reference labeling antibodies, primary antibodies and secondary antibodies of immunological experiments, etc.; favorite products of cell research users, from Dyes and kits for detecting the state of cells, kits for extracting organelles, staining and tracking of cell substructures and cell metabolism detection products, to cytokine and protein detection kits for cell culture, only to help your research career !
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.
BioAdvisers said on Biotech Advisers
Immunofluorescence (IF) how to make pictures more colorful
Junior: Brother, brother, come out soon
Senior: What should I do to my sister and sister, little sister
Junior: The fluorescence of IF I made with NeuN antibody is not clear, is it the reason for the secondary antibody?
Junior:
Senior: NeuN is a mature nuclear antigen, which focuses on nuclear localization. There are many reasons for the blurring of fluorescence. Judging from the results, the causes of antibodies can be eliminated.
Junior: We generally think that it is the cause of antibodies if the experiment is unsuccessful. Why is this not the cause of antibodies this time?
Senior: That’s for sure! Let me tell you slowly.
Each picture of immunofluorescence is a work of art. If you are a senior player of experimental immunohistochemistry, you can take a picture of it like this:
It is from http://www.fansbio.com/content/?400.html
It is from http://www.dxy.cn/bbs/thread/38628725?sf=2&dn=5#38628725
https://www.biofavor.cn/index-show-tid-200.html
http://www.rolesbio.com/product/5069.html
Various tissue shapes, cell structures, and protein shapes are scattered and organically combined to form a colorful and exquisite fluorescent world, uncovering the mysteries of scientific research one after another. It’s really beautiful (du).
So, how to operate in order to obtain a series of perfect immunofluorescence results like a senior player?
Today, I will take you to take a closer look at the immunofluorescence experiment~
Experimental principle:
Immunofluorescence (immunofluorescence technic): Coons equal to the first use of fluorescein for labeling in 1941 and achieved success. This technique of labeling antibodies with fluorescent substances for antigen localization is called fluorescent antibody technology (fluorescent antibody technique). The method of tracing or checking the corresponding antigen with fluorescent antibodies is called the fluorescent antibody method; the method of tracing or checking the corresponding antibodies with known fluorescent antigen labels is called the fluorescent antigen method. These two methods are collectively referred to as immunofluorescence technology, because fluorescent pigments can not only bind to antibody globulin for detecting or localizing various antigens, but also bind to other proteins for detecting or localizing antibodies, but fluorescent antigens are used in actual work. Technology is rarely used, so people are accustomed to call it fluorescent antibody technology, or immunofluorescence technology. The fluorescent antibody method is more commonly used. Immunofluorescence technology to display and examine the antigen or hapten material in cells or tissues is called immunofluorescence cell (or tissue) chemical technology.
Steps:
One. Sample processing
Ⅰ Cell slide
In the cell culture well plate, the slides that have climbed the cells are rinsed with PBS 3 times, 5 min each time.
Fix in 4% paraformaldehyde or other fixing solution at room temperature for 15 minutes, wash three times with PBST for 5 minutes each time (fixation method is not unique, depending on the total cell type).
Ⅱ Frozen section
Frozen tissues of fixed tissues were equilibrated and rewarmed at room temperature for 30 minutes;
Wash three times with PBST, 5min each time;
Ø Paraffin section
Dewaxing Hydration
Soak in xylene Ⅰ cylinder for 15min;
Immersed in xylene II tank for 15min;
Soak in the No. 1 cylinder of absolute ethanol for 5min;
Soak in absolute ethanol No. 2 cylinder for 5min;
Soak in 95% ethanol for 5min;
Soak in 80% ethanol for 5min;
Soak in 60% ethanol for 5min;
Wash with deionized water 3 times, 5min each time.
Here, cylinders I and II refer to different containers, but the contents are the same.
Antigen retrieval
Take an appropriate amount of sodium citrate repair solution (pH 6.0) or Tris-EDTA repair solution (pH 9.0) in a glass beaker (the repair solution can not pass the slice holder), after the repair solution is boiled, put the slices in it, Close the lid and continue to boil the repair solution for 15min. Cool naturally in a fume kitchen.
two. Antibody recognition and fluorescence photography
Wash the slides 3 times with PBST, 3 minutes each time, absorb the PBST with absorbent paper, add normal goat serum onto the slides, and seal at room temperature for 30 minutes;
Absorb the blocking solution with absorbent paper, do not wash, add enough diluted primary antibody dropwise to each slide and put it in the wet box, incubate at room temperature for 2-3h (or incubate overnight at four degrees); secondary antibody incubate at room temperature Overnight at 1h or 4°C. Note: Starting with the addition of fluorescent primary antibodies, all subsequent steps should be carried out in a dark place.
Counterstaining nucleus: add DAPI and incubate in the dark for 5min to stain the specimen, PBST 5min × 4 times to wash away excess DAPI; 4. Absorb excess liquid on the slide with absorbent paper and quench with anti-fluorescence The mounting solution of the agent is mounted, and then observed and collected under a fluorescence microscope.
Troubleshooting FAQ
What should I do with irregularly shaped bright spots captured by immunofluorescence microscopy?
Irregular and more bright spots are generally due to impurities or dust on the surface of the slide material, which can be avoided by adjusting the focal plane to the tissue.
What is the method of channel switching during the photo?
First find the target position under white light, and adjust to the clearest focal length, and then switch to the fluorescent channel to shoot.
How to avoid dye fading?
In order to reduce the degree of fading of some samples, it is recommended to use neutral density filters in the light path before the light reaches the excitation filter, thereby reducing the intensity of the excitation light. In other cases, the fading effect can be reduced by changing the pH of the mounting medium or by using anti-bleaching agents (a few more important agents are listed in Table 2). For digital imaging, microphotography or simple visual observation, changing the field of view quickly can also avoid the fading effect.
What is the reason why the organization is clear in the process of taking pictures?
In the process of making tissue sections, there are some areas that are not firmly attached to the slides, and there is a tendency to take off the slide, resulting in the sample not on a focal plane, and a blurred image under the microscope.
No staining of tissue cells
One case is experimental operation problems, including inaccurate experimental operation methods, stale experimental materials, antibody species and reactivity selection, and antibody quality problems. These can be eliminated by setting up positive and negative control experiments. The second situation is the protein expression problem. Like the “WB experiment without bands” mentioned in the previous issue, the IF experiment preparation stage also needs to consult a lot of data to initially confirm the spatiotemporal expression profile of the target protein (including expression amount, expression site, and stimulation conditions Wait).
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.
Bioadvisers shared on Biotech Advisers
Immunofluorescence (IF) how to make pictures more colorful
Junior: Brother, brother, come out soon
Senior: What should I do to my sister and sister, little sister
Junior: The fluorescence of IF I made with NeuN antibody is not clear, is it the reason for the secondary antibody?
Junior:
Senior: NeuN is a mature nuclear antigen, which focuses on nuclear localization. There are many reasons for the blurring of fluorescence. Judging from the results, the causes of antibodies can be eliminated.
Junior: We generally think that it is the cause of antibodies if the experiment is unsuccessful. Why is this not the cause of antibodies this time?
Senior: That’s for sure! Let me tell you slowly.
Each picture of immunofluorescence is a work of art. If you are a senior player of experimental immunohistochemistry, you can take a picture of it like this:
It is from http://www.fansbio.com/content/?400.html
It is from http://www.dxy.cn/bbs/thread/38628725?sf=2&dn=5#38628725
https://www.biofavor.cn/index-show-tid-200.html
http://www.rolesbio.com/product/5069.html
Various tissue shapes, cell structures, and protein shapes are scattered and organically combined to form a colorful and exquisite fluorescent world, uncovering the mysteries of scientific research one after another. It’s really beautiful (du).
So, how to operate in order to obtain a series of perfect immunofluorescence results like a senior player?
Today, I will take you to take a closer look at the immunofluorescence experiment~
Experimental principle:
Immunofluorescence (immunofluorescence technic): Coons equal to the first use of fluorescein for labeling in 1941 and achieved success. This technique of labeling antibodies with fluorescent substances for antigen localization is called fluorescent antibody technology (fluorescent antibody technique). The method of tracing or checking the corresponding antigen with fluorescent antibodies is called the fluorescent antibody method; the method of tracing or checking the corresponding antibodies with known fluorescent antigen labels is called the fluorescent antigen method. These two methods are collectively referred to as immunofluorescence technology, because fluorescent pigments can not only bind to antibody globulin for detecting or localizing various antigens, but also bind to other proteins for detecting or localizing antibodies, but fluorescent antigens are used in actual work. Technology is rarely used, so people are accustomed to call it fluorescent antibody technology, or immunofluorescence technology. The fluorescent antibody method is more commonly used. Immunofluorescence technology to display and examine the antigen or hapten material in cells or tissues is called immunofluorescence cell (or tissue) chemical technology.
Steps:
One. Sample processing
Ⅰ Cell slide
In the cell culture well plate, the slides that have climbed the cells are rinsed with PBS 3 times, 5 min each time.
Fix in 4% paraformaldehyde or other fixing solution at room temperature for 15 minutes, wash three times with PBST for 5 minutes each time (fixation method is not unique, depending on the total cell type).
Ⅱ Frozen section
Frozen tissues of fixed tissues were equilibrated and rewarmed at room temperature for 30 minutes;
Wash three times with PBST, 5min each time;
Ø Paraffin section
Dewaxing Hydration
Soak in xylene Ⅰ cylinder for 15min;
Immersed in xylene II tank for 15min;
Soak in the No. 1 cylinder of absolute ethanol for 5min;
Soak in absolute ethanol No. 2 cylinder for 5min;
Soak in 95% ethanol for 5min;
Soak in 80% ethanol for 5min;
Soak in 60% ethanol for 5min;
Wash with deionized water 3 times, 5min each time.
Here, cylinders I and II refer to different containers, but the contents are the same.
Antigen retrieval
Take an appropriate amount of sodium citrate repair solution (pH 6.0) or Tris-EDTA repair solution (pH 9.0) in a glass beaker (the repair solution can not pass the slice holder), after the repair solution is boiled, put the slices in it, Close the lid and continue to boil the repair solution for 15min. Cool naturally in a fume kitchen.
two. Antibody recognition and fluorescence photography
Wash the slides 3 times with PBST, 3 minutes each time, absorb the PBST with absorbent paper, add normal goat serum onto the slides, and seal at room temperature for 30 minutes;
Absorb the blocking solution with absorbent paper, do not wash, add enough diluted primary antibody dropwise to each slide and put it in the wet box, incubate at room temperature for 2-3h (or incubate overnight at four degrees); secondary antibody incubate at room temperature Overnight at 1h or 4°C. Note: Starting with the addition of fluorescent primary antibodies, all subsequent steps should be carried out in a dark place.
Counterstaining nucleus: add DAPI and incubate in the dark for 5min to stain the specimen, PBST 5min × 4 times to wash away excess DAPI; 4. Absorb excess liquid on the slide with absorbent paper and quench with anti-fluorescence The mounting solution of the agent is mounted, and then observed and collected under a fluorescence microscope.
Troubleshooting FAQ
What should I do with irregularly shaped bright spots captured by immunofluorescence microscopy?
Irregular and more bright spots are generally due to impurities or dust on the surface of the slide material, which can be avoided by adjusting the focal plane to the tissue.
What is the method of channel switching during the photo?
First find the target position under white light, and adjust to the clearest focal length, and then switch to the fluorescent channel to shoot.
How to avoid dye fading?
In order to reduce the degree of fading of some samples, it is recommended to use neutral density filters in the light path before the light reaches the excitation filter, thereby reducing the intensity of the excitation light. In other cases, the fading effect can be reduced by changing the pH of the mounting medium or by using anti-bleaching agents (a few more important agents are listed in Table 2). For digital imaging, microphotography or simple visual observation, changing the field of view quickly can also avoid the fading effect.
What is the reason why the organization is clear in the process of taking pictures?
In the process of making tissue sections, there are some areas that are not firmly attached to the slides, and there is a tendency to take off the slide, resulting in the sample not on a focal plane, and a blurred image under the microscope.
No staining of tissue cells
One case is experimental operation problems, including inaccurate experimental operation methods, stale experimental materials, antibody species and reactivity selection, and antibody quality problems. These can be eliminated by setting up positive and negative control experiments. The second situation is the protein expression problem. Like the “WB experiment without bands” mentioned in the previous issue, the IF experiment preparation stage also needs to consult a lot of data to initially confirm the spatiotemporal expression profile of the target protein (including expression amount, expression site, and stimulation conditions Wait).
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.
Bioadvisers shared on Biotech Advisers
Immunofluorescence (IF) how to make pictures more colorful
Junior: Brother, brother, come out soon
Senior: What should I do to my sister and sister, little sister
Junior: The fluorescence of IF I made with NeuN antibody is not clear, is it the reason for the secondary antibody?
Junior:
Senior: NeuN is a mature nuclear antigen, which focuses on nuclear localization. There are many reasons for the blurring of fluorescence. Judging from the results, the causes of antibodies can be eliminated.
Junior: We generally think that it is the cause of antibodies if the experiment is unsuccessful. Why is this not the cause of antibodies this time?
Senior: That’s for sure! Let me tell you slowly.
Each picture of immunofluorescence is a work of art. If you are a senior player of experimental immunohistochemistry, you can take a picture of it like this:
It is from http://www.fansbio.com/content/?400.html
It is from http://www.dxy.cn/bbs/thread/38628725?sf=2&dn=5#38628725
https://www.biofavor.cn/index-show-tid-200.html
http://www.rolesbio.com/product/5069.html
Various tissue shapes, cell structures, and protein shapes are scattered and organically combined to form a colorful and exquisite fluorescent world, uncovering the mysteries of scientific research one after another. It’s really beautiful (du).
So, how to operate in order to obtain a series of perfect immunofluorescence results like a senior player?
Today, I will take you to take a closer look at the immunofluorescence experiment~
Experimental principle:
Immunofluorescence (immunofluorescence technic): Coons equal to the first use of fluorescein for labeling in 1941 and achieved success. This technique of labeling antibodies with fluorescent substances for antigen localization is called fluorescent antibody technology (fluorescent antibody technique). The method of tracing or checking the corresponding antigen with fluorescent antibodies is called the fluorescent antibody method; the method of tracing or checking the corresponding antibodies with known fluorescent antigen labels is called the fluorescent antigen method. These two methods are collectively referred to as immunofluorescence technology, because fluorescent pigments can not only bind to antibody globulin for detecting or localizing various antigens, but also bind to other proteins for detecting or localizing antibodies, but fluorescent antigens are used in actual work. Technology is rarely used, so people are accustomed to call it fluorescent antibody technology, or immunofluorescence technology. The fluorescent antibody method is more commonly used. Immunofluorescence technology to display and examine the antigen or hapten material in cells or tissues is called immunofluorescence cell (or tissue) chemical technology.
Steps:
One. Sample processing
Ⅰ Cell slide
In the cell culture well plate, the slides that have climbed the cells are rinsed with PBS 3 times, 5 min each time.
Fix in 4% paraformaldehyde or other fixing solution at room temperature for 15 minutes, wash three times with PBST for 5 minutes each time (fixation method is not unique, depending on the total cell type).
Ⅱ Frozen section
Frozen tissues of fixed tissues were equilibrated and rewarmed at room temperature for 30 minutes;
Wash three times with PBST, 5min each time;
Ø Paraffin section
Dewaxing Hydration
Soak in xylene Ⅰ cylinder for 15min;
Immersed in xylene II tank for 15min;
Soak in the No. 1 cylinder of absolute ethanol for 5min;
Soak in absolute ethanol No. 2 cylinder for 5min;
Soak in 95% ethanol for 5min;
Soak in 80% ethanol for 5min;
Soak in 60% ethanol for 5min;
Wash with deionized water 3 times, 5min each time.
Here, cylinders I and II refer to different containers, but the contents are the same.
Antigen retrieval
Take an appropriate amount of sodium citrate repair solution (pH 6.0) or Tris-EDTA repair solution (pH 9.0) in a glass beaker (the repair solution can not pass the slice holder), after the repair solution is boiled, put the slices in it, Close the lid and continue to boil the repair solution for 15min. Cool naturally in a fume kitchen.
two. Antibody recognition and fluorescence photography
Wash the slides 3 times with PBST, 3 minutes each time, absorb the PBST with absorbent paper, add normal goat serum onto the slides, and seal at room temperature for 30 minutes;
Absorb the blocking solution with absorbent paper, do not wash, add enough diluted primary antibody dropwise to each slide and put it in the wet box, incubate at room temperature for 2-3h (or incubate overnight at four degrees); secondary antibody incubate at room temperature Overnight at 1h or 4°C. Note: Starting with the addition of fluorescent primary antibodies, all subsequent steps should be carried out in a dark place.
Counterstaining nucleus: add DAPI and incubate in the dark for 5min to stain the specimen, PBST 5min × 4 times to wash away excess DAPI; 4. Absorb excess liquid on the slide with absorbent paper and quench with anti-fluorescence The mounting solution of the agent is mounted, and then observed and collected under a fluorescence microscope.
Troubleshooting FAQ
What should I do with irregularly shaped bright spots captured by immunofluorescence microscopy?
Irregular and more bright spots are generally due to impurities or dust on the surface of the slide material, which can be avoided by adjusting the focal plane to the tissue.
What is the method of channel switching during the photo?
First find the target position under white light, and adjust to the clearest focal length, and then switch to the fluorescent channel to shoot.
How to avoid dye fading?
In order to reduce the degree of fading of some samples, it is recommended to use neutral density filters in the light path before the light reaches the excitation filter, thereby reducing the intensity of the excitation light. In other cases, the fading effect can be reduced by changing the pH of the mounting medium or by using anti-bleaching agents (a few more important agents are listed in Table 2). For digital imaging, microphotography or simple visual observation, changing the field of view quickly can also avoid the fading effect.
What is the reason why the organization is clear in the process of taking pictures?
In the process of making tissue sections, there are some areas that are not firmly attached to the slides, and there is a tendency to take off the slide, resulting in the sample not on a focal plane, and a blurred image under the microscope.
No staining of tissue cells
One case is experimental operation problems, including inaccurate experimental operation methods, stale experimental materials, antibody species and reactivity selection, and antibody quality problems. These can be eliminated by setting up positive and negative control experiments. The second situation is the protein expression problem. Like the “WB experiment without bands” mentioned in the previous issue, the IF experiment preparation stage also needs to consult a lot of data to initially confirm the spatiotemporal expression profile of the target protein (including expression amount, expression site, and stimulation conditions Wait).
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.
