Visualization of multiple protein bands on the same nitrocellulose membrane by double immunoblotting

Anal Biochem. 1988 Nov 15;175(1):191-5. doi: 10.1016/0003-2697(88)90377-6.

Abstract

A method has been developed which allows the simultaneous immunodetection of more than one type of protein on the same nitrocellulose membrane. This procedure does not require special labeling of samples or elution of antibodies from the membrane as do the alternatives cited in the literature (1,2). Proteins are separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred to the membrane before specific immunostaining with either peroxidase/4-chloro-1-naphthol or immunogold/silver staining. Antigen identity is visually determined by the formation of different-colored precipitates on the membrane. This innovation in protein blotting offers a savings in time and reagents as well as permitting identification of closely spaced bands with certainty.

MeSH terms

  • Bacterial Proteins / isolation & purification
  • Blotting, Western / methods*
  • Collodion
  • Gold
  • Growth Hormone / analogs & derivatives
  • Growth Hormone / isolation & purification
  • Horseradish Peroxidase
  • Human Growth Hormone
  • Naphthols
  • Proteins / isolation & purification*
  • Silver
  • Staining and Labeling

Substances

  • Bacterial Proteins
  • Naphthols
  • Proteins
  • Human Growth Hormone
  • Silver
  • 4-chloro-1-naphthol
  • Gold
  • Growth Hormone
  • Collodion
  • Horseradish Peroxidase