Open AccessDetection of Proteins on Blot Membranes
Author(s) -
Goldman Aaron,
Harper Sandra,
Speicher David W.
Publication year - 2016
Publication title -
current protocols in protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.409
H-Index - 32
eISSN - 1934-3663
pISSN - 1934-3655
DOI - 10.1002/cpps.15
Subject(s) - membrane , western blot , chemistry , microbiology and biotechnology , biochemistry , computational biology , biology , gene
Staining of blot membranes enables the visualization of bound proteins. Proteins are usually transferred to blot membranes by electroblotting, by direct spotting of protein solutions, or by contact blots. Staining allows the efficiency of transfer to the membrane to be monitored. This unit describes protocols for staining proteins after electroblotting from polyacrylamide gels to blot membranes such as polyvinylidene difluoride (PVDF), nitrocellulose, or nylon membranes. The same methods can be used if proteins are directly spotted, either manually or using robotics. Protocols are included for seven general protein stains (amido black, Coomassie blue, Ponceau S, colloidal gold, colloidal silver, India ink, and MemCode) and three fluorescent protein stains (fluorescamine, IAEDANS, and SYPRO Ruby). Also included is an in‐depth discussion of the different blot membrane types and the compatibility of different protein stains with downstream applications, such as immunoblotting or N‐terminal Edman sequencing. © 2016 by John Wiley & Sons, Inc.