PMID: 7535985Jan 1, 1995Paper

Double staining of coomassie blue-stained polyacrylamide gels by imidazole-sodium dodecyl sulfate-zinc reverse staining: sensitive detection of coomassie blue-undetected proteins

Analytical Biochemistry
C Fernandez-PatronL Castellanos

Abstract

The sensitivity, simplicity, and relative rapidity of Coomassie blue staining have made this technique the method of choice for routine detection and quantitative analysis of gel electrophoresis-separated protein bands in many applications. To extend the usefulness of this technique, we have developed a new double-staining method for visualizing SDS-PAGE-separated protein bands that were undetected by Coomassie blue staining of the gel. Coomassie blue-stained gels are washed in distilled water (15 min, two times) and then subjected to imidazole-zinc reverse staining. As a result of the method, a homogeneous white-stained background is generated and two types of protein bands can be observed: (a) typical Coomassie blue-stained bands, which appear superposed on larger transparent bands; and (b) reverse-stained (transparent) bands, which were previously undetected by the Coomassie blue staining. The method is rapid, simple, and reproducible and double-staining gels can be kept in distilled water for months without loss of the protein pattern. The overall sensitivity is high (e.g., 1.6 ng for recombinant streptokinase, 47 kDa) over a wide range of protein molecular weights (10 to 100 kDa) and independent of the degree of Coomassie ...Continue Reading

Citations

Mar 6, 2007·Nature·Jessica ReinekeChristoph von Eichel-Streiber
Jul 31, 2007·Rapid Communications in Mass Spectrometry : RCM·Akihiro ItoShigemi Norioka
Sep 1, 2011·Mass Spectrometry Reviews·Peihong ZhuJohn G Marshall
Apr 30, 2003·Rapid Communications in Mass Spectrometry : RCM·Hiroyuki KatayamaTakeshi Nagasu
May 13, 2010·Biological & Pharmaceutical Bulletin·Masae OhnoMasayoshi Imagawa
Jun 21, 2011·Journal of Chemical Biology·Victoria J GauciJens R Coorssen
Dec 31, 2014·Microbial Cell Factories·Bertrand RaynalPatrick England
Jan 30, 2008·Current Protocols in Cell Biology·E C Dell'Angelica, J S Bonifacino
Jan 30, 2008·Current Protocols in Cell Biology·Adam BrymoraPhillip J Robinson
Apr 5, 2007·Nature Protocols·Lila Castellanos-Serra, Eugenio Hardy
Feb 22, 2021·The Journal of Biological Chemistry·Ku-Feng LinWilliam M Clemons
Feb 6, 2020·Methods : a Companion to Methods in Enzymology·Amanda L WongJames J Pesavento
Apr 21, 2006·International Journal of Biological Macromolecules·Melanie M HoeferHarald Illges

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