UNIT 8.10B Detection of Proteins on Blot Transfer Membranes

  1. Joachim Sasse1,
  2. Sean R. Gallagher2

Published Online: 1 NOV 2008

DOI: 10.1002/0471142735.im0810bs83

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Sasse, J. and Gallagher, S. R. 2008. Detection of Proteins on Blot Transfer Membranes. Current Protocols in Immunology. 83:IV:8.10B:8.10B.1–8.10B.6.

Author Information

  1. 1

    Shriners Hospital for Crippled Children, Tampa, Florida

  2. 2

    UVP, Inc., Upland, California

Publication History

  1. Published Online: 1 NOV 2008
  2. Published Print: NOV 2008


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. Curr. Protoc. Immunol. 83:8.10B.1-8.10B.6. © 2008 by John Wiley & Sons, Inc.


  • nylon;
  • nitrocellulose;
  • PVDF;
  • protein/blotting;
  • stain;
  • total protein;
  • fluorescence