Western blotting

Methods. 2006 Apr;38(4):283-93. doi: 10.1016/j.ymeth.2005.11.007.

Abstract

Western blotting (protein blotting or immunoblotting) is a powerful and important procedure for the immunodetection of proteins post-electrophoresis, particularly proteins that are of low abundance. Since the inception of the protocol for protein transfer from an electrophoresed gel to a membrane in 1979, protein blotting has evolved greatly. The scientific community is now confronted with a variety of ways and means to carry out this transfer. This review describes the various procedures that have been used to transfer proteins from a gel to a membrane based on the principles of simple diffusion, vacuum-assisted solvent flow and electrophoretic elution. Finally, a brief description of methods generally used to detect antigens on blots is also described.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens / chemistry
  • Biotinylation
  • Blotting, Western / instrumentation*
  • Blotting, Western / methods*
  • Collodion / chemistry
  • Diffusion
  • Electrophoresis
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Immunoblotting
  • Membranes, Artificial
  • Proteins / analysis*
  • Proteins / chemistry
  • Solvents

Substances

  • Antigens
  • Membranes, Artificial
  • Proteins
  • Solvents
  • Collodion