The Western Blot

Methods Mol Biol. 2015:1318:87-96. doi: 10.1007/978-1-4939-2742-5_9.

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 semiquantitative 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.

MeSH terms

  • Animals
  • Antibodies / chemistry*
  • Blotting, Western / instrumentation
  • Blotting, Western / methods*
  • Collodion
  • Electrophoresis, Polyacrylamide Gel / instrumentation
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Humans
  • Molecular Weight
  • Polyvinyls
  • Proteins / isolation & purification*
  • Sensitivity and Specificity

Substances

  • Antibodies
  • Polyvinyls
  • Proteins
  • polyvinylidene fluoride
  • Collodion