Location: Produce Safety and Microbiology ResearchTitle: The western blot
|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.