Detection of proteins on blot membranes

Curr Protoc Protein Sci. 2001 May:Chapter 10:Unit 10.8. doi: 10.1002/0471140864.ps1008s00.

Abstract

Staining of blot transfer membranes permits visualization of proteins and allows the extent of transfer to be monitored. In the protocols described in this unit, proteins are stained after electroblotting from one-dimensional or two-dimensional polyacrylamide gels to blot membranes such as polyvinylidene difluoride (PVDF), nitrocellulose, or nylon membranes. Protocols are provided for the use of six general protein stains: amido black, Coomassie blue, Ponceau S, colloidal gold, colloidal silver, and India ink. In addition, the fluorescent stains fluorescamine and IAEDANS, which covalently react with bound proteins, are described. Approximate detection limits for each nonfluorescent stain are indicated along with membrane compatibilities.

MeSH terms

  • Amido Black
  • Azo Compounds
  • Blotting, Western / methods*
  • Carbon
  • Fluorescamine
  • Gold Colloid
  • Membranes, Artificial*
  • Naphthalenesulfonates
  • Proteins / analysis*
  • Rosaniline Dyes
  • Staining and Labeling

Substances

  • Azo Compounds
  • Gold Colloid
  • Membranes, Artificial
  • Naphthalenesulfonates
  • Proteins
  • Rosaniline Dyes
  • chinese ink
  • Fluorescamine
  • 1,5-I-AEDANS
  • Carbon
  • Coomassie blue
  • Amido Black
  • ponceau S