Detection of Proteins on Blot Membranes
Open Access
- 1 November 2016
- journal article
- unit
- Published by Wiley in Current Protocols in Protein Science
- Vol. 86 (1), 10.8.1-10.8.11
- https://doi.org/10.1002/cpps.15
Abstract
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.This publication has 14 references indexed in Scilit:
- Analysis of trypsin inhibition activity in human plasma proteins after separation by non-denaturing two-dimensional electrophoresisClinica Chimica Acta; International Journal of Clinical Chemistry, 2013
- One- and Two-Dimensional Blue Native-PAGE and Immunodetection of Low-Abundance Chloroplast Membrane Protein ComplexesMethods in molecular biology (Clifton, N.J.), 2011
- A high-affinity reversible protein stain for Western blotsAnalytical Biochemistry, 2004
- A Luminescent Ruthenium Complex for Ultrasensitive Detection of Proteins Immobilized on Membrane SupportsAnalytical Biochemistry, 1999
- Microsequence analysis of electroblotted proteinsAnalytical Biochemistry, 1992
- MICROSEQUENCING WITH PVDF MEMBRANES: EFFICIENT ELECTROBLOTTING, DIRECT PROTEIN ADSORPTION AND SEQUENCER PROGRAM MODIFICATIONSPublished by Elsevier BV ,1989
- Fluorescent labeling of nitrocellulose-bound proteins at the nanogram level without changes in immunoreactivityAnalytical Biochemistry, 1988
- Alkali enhancement of protein staining on nitrocelluloseElectrophoresis, 1986
- Sensitive colloidal metal (gold or silver) staining of protein blots on nitrocellulose membranesAnalytical Biochemistry, 1985
- India ink staining of proteins on nitrocellulose paperAnalytical Biochemistry, 1983