Detection of Proteins on Polyacrylamide Gels Using Calconcarboxylic Acid

✍ Scribed by H.Y. Hong; J.K. Choi; G.S. Yoo

• Publisher: Elsevier Science
• Year: 1993
• Tongue: English
• Weight: 418 KB
• Volume: 214
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1993.1461

✦ Synopsis

We describe here a protein staining method in polyacrylamide gels with a new staining dye, 1-(2-hydroxy4-sulfo-1-naphthylazo)-2-hydroxy-3-naphthoic acid (calconcarboxylic acid, NN). This method can be performed by both simultaneous and postelectrophoretic staining techniques. Simultaneous staining using (0.01 %) of (\mathrm{NN}) in upper reservoir buffer eliminates the poststaining step, and thus enables detection of the proteins more rapidly and simply. In poststaining, proteins can be stained by a (30-\mathrm{min}) incubation of a polyacrylamide gel in (40 %) methanol (/ 7 %) acetic acid solution of (0.05 % \mathrm{NN}). These techniques produced protein staining patterns identical to the ones obtained by the conventional poststaining with Coomassie blue R-250 (CB). (\mathrm{NN}) staining can detect as little as (10 \mathrm{ng}) of bovine serum albumin by poststaining and (25 \mathrm{ng}) by simultaneous staining, compared to (50 \mathrm{ng}) detectable by (\mathrm{CB}) poststaining. In comparing the relationship between band intensity and amount of protein, NN staining gave better linearity than CB staining. (c) 1993 Academic Press, Ine.