Western Blot Introduction
Western Blot is a technique used to detect specific proteins from samples like cell extracts or tissue homogenates. This technique is firstly developed by W. Neal Burnette, and the name “Western Blot” is a play on the name Southern Blot, which is named after its inventor Edwin Southern.The western blot (sometimes called the protein immunoblot) is a widely used analytical technique used to detect specific proteins in the given sample of tissue homogenate or extract. The specificity of the antibody-antigen interaction enables a target protein to be identified in the midst of a complex protein mixture. Western blotting can produce qualitative and semiquantitative data about that protein.It uses gel electrophoresis to separate native or denatured proteins by the length of the polypeptide (denaturing conditions) or by the 3-D structure of the protein (native/ non-denaturing conditions). The proteins are then transferred to a membrane (typically nitrocellulose or PVDF), where they are probed (detected) using antibodies specific to the target protein.
References:
E.M. Southern, Detection of specific sequences among DNA fragments separated by gel electrophoresis, Journal of Molecular Biology, Volume 98, Issue 3, 5 November 1975, Pages 503-517, ISSN 0022-2836, DOI: 10.1016/S0022-2836(75)80083-0.
W. Neal Burnette, 'Western Blotting': Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A, Analytical Biochemistry, Volume 112, Issue 2, April 1981, Pages 195-203, ISSN 0003-2697, DOI: 10.1016/0003-2697(81)90281-5.