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ARS Home » Pacific West Area » Albany, California » Western Regional Research Center » Produce Safety and Microbiology Research » Research » Publications at this Location » Publication #305667

Research Project: Immunodiagnostics to Detect Prions and Other Important Animal Pathogens

Location: Produce Safety and Microbiology Research

Title: The western blot

Author
item Hnasko, Robert
item HNASKO, THOMAS - University Of California

Submitted to: Methods in Molecular Biology
Publication Type: Book / Chapter
Publication Acceptance Date: 4/4/2015
Publication Date: 7/14/2015
Citation: Hnasko, R.M., Hnasko, T.S. 2015. The western blot. In: Hnasko, Robert, editor. ELISA: Methods and protocols. Methods in Molecular Biology. 1st edition. New York, NY:Humana Press. p. 87-96. doi: 10.1007/978-1-4939-2742-5.

Interpretive Summary: This manuscript provides details on how to use an antibody to conduct a Western blot immunoassay for the detection of a target antigen from a diagnostic sample and obtain its mass.

Technical Abstract: Western blotting is a technique that involves the separation of proteins by gel electrophoresis, their blotting or transfer to a membrane, and selective immunodetection of an immobilized antigen. This is an important and routine method for protein analysis that depends on the specificity of antibody-antigen interaction and is useful for the qualitative or semi-quantitative identification of specific proteins and their molecular weight from a complex mixture. This chapter will outline the requisite steps including: gel electrophoresis of a protein sample, transfer of protein from a gel to a membrane support, and immunodetection of a target antigen.