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Immunoblotting and Immunodetection
Current Protocols in Immunology, 2016AbstractImmunoblotting (western blotting) is used to identify specific antigens recognized by polyclonal or monoclonal antibodies. This unit provides protocols for all steps, starting with solubilization of the protein samples, usually by means of SDS and reducing agents.
Duojiao, Ni, Peng, Xu, Sean, Gallagher
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2009
Grid-Immunoblotting is a fast, simple, and efficient method for simultaneously testing multiple allergens utilizing small amount of antibody.
Olga, Yeliosof, R Hal, Scofield
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Grid-Immunoblotting is a fast, simple, and efficient method for simultaneously testing multiple allergens utilizing small amount of antibody.
Olga, Yeliosof, R Hal, Scofield
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Current Protocols in Protein Science, 1996
AbstractImmunoblotting (often referred to as western blotting) is used to identify specific antigens recognized by polyclonal or monoclonal antibodies. Following electrophoretic separation of proteins and transfer from the gel to an appropriate membrane, the immobilized proteins are probed with specific antibodies to identify and quantitate any ...
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AbstractImmunoblotting (often referred to as western blotting) is used to identify specific antigens recognized by polyclonal or monoclonal antibodies. Following electrophoretic separation of proteins and transfer from the gel to an appropriate membrane, the immobilized proteins are probed with specific antibodies to identify and quantitate any ...
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Immunoblotting for Calpastatin Expression
2019Immunoblotting is a procedure routinely used to analyze calpastatin expression. However, immunoblotting alone may not be adequate for this task, since calpastatin isoforms can vary by tissue, can be modified by partial digestion, and can undergo posttranslational modifications.
Averna, Monica, De Tullio, Roberta
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Diagnosis of Paragonimiasis by Immunoblot
The American Journal of Tropical Medicine and Hygiene, 1988A sensitive and specific immunoblot assay was used to rapidly and accurately diagnose paragonimiasis. The immunoreactivity of a complex Paragonimus westermani Chaffee antigen was evaluated by SDS-PAGE and Western blot analysis. Initial probing with pooled human serum from proven Paragonimus infections revealed many bands, including a significant ...
S B, Slemenda +3 more
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Preparing Immunoprecipitations for Immunoblotting
Cold Spring Harbor Protocols, 2020Immunoprecipitated proteins can be readily analyzed by immunoblotting. Proteins can be efficiently eluted from the Protein A or similar beads by addition of the SDS-PAGE sample loading buffer and heating at 95°C. This elution procedure will also remove the capturing antibody from the beads unless the antibody was cross-linked to the beads ...
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Detection of an Antigen on an Immunoblot
Cold Spring Harbor Protocols, 2019Detection of the antigen on an immunoblot can be achieved by either enzyme-based detection systems or using detection reagents labeled with fluorochromes. Two major types of enzyme-based detection systems are used in immunoblotting based on either horseradish peroxidase (HRP)–or alkaline phosphatase (AP)–coupled antibodies, and a range of soluble ...
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Stripping of the Immunoblot for Reprobing
Cold Spring Harbor Protocols, 2020For most immunoblots developed with chemiluminescence or with fluorochrome-based detection systems, it is possible to remove the primary and secondary antibodies from the membrane without affecting the bound antigen. This allows you to reuse the membrane for detection of another protein antigen.
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Protein Blotting and Immunoblotting
2000Immunoblotting was first devised by Towbin et al. (1979) to exploit the specificity of the reaction between antibodies and proteins transferred from SDS- polyacrylamide electrophoretic gels onto nitrocellulose. Immunoreactive bands were evidenced by labelled “second antibody” or protein A.
BINI, LUCA +5 more
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Visualization of the Insulin Receptor by Immunoblotting
Diabetes, 1984Insulin receptors from rat hepatoma cells (Fao) and human placenta were partially purified by detergent solu-bilization and lectin purification. The insulin receptor preparations were subjected to sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis under reducing or nonreducing conditions.
R, Maron +3 more
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