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Updated: Jun 27, 2026

Immunoblot Analysis
16:01

Immunoblot Analysis

Published on: June 20, 2008

Immunoblot analysis

Sean Gallagher1, Deb Chakavarti

  • 1UVP, LLC, USA.

Journal of Visualized Experiments : Jove
|December 11, 2008
PubMed

Immunoblotting (western blotting) is a rapid and sensitive assay for the detection and characterization of proteins that works by exploiting the specificity inherent in antigen-antibody recognition. It involves the solubilization and electrophoretic separation of proteins, glycoproteins, or lipopolysaccharides by gel electrophoresis, followed by quantitative transfer and irreversible binding to nitrocellulose, PVDF, or nylon. The immunoblotting technique has been useful in identifying specific antigens recognized by polyclonal or monoclonal antibodies and is highly sensitive (1 ng of antigen can be detected). This unit provides protocols for protein separation, blotting proteins onto membranes, immunoprobing, and visualization using chromogenic or chemiluminescent substrates.

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