Immunoblot Analysis | Zendy

Sean R. Gallagher | Zendy; Deb Chakavarti | Zendy

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Immunoblot Analysis

Author(s) -

Sean R. Gallagher,

Deb Chakavarti

Publication year - 2008

Publication title -

journal of visualized experiments

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.596

H-Index - 91

ISSN - 1940-087X

DOI - 10.3791/759

Subject(s) - polyclonal antibodies , nitrocellulose , blot , microbiology and biotechnology , glycoprotein , antigen , monoclonal antibody , chromogenic , chemistry , electroblotting , gel electrophoresis , antibody , biochemistry , biology , membrane , chromatography , immunology , gene

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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