A simple method for staining of proteins simultaneously on sodium dodecyl sulfate (SDS)–polyacrylamide gels and nitrocellulose membranes by Alta during Western blot analysis is described. A 5% solution of Alta, a commercially available cosmetic preparation, is added in the upper tank buffer during electrophoresis. On completion of electrophoresis, the gel is washed in distilled water, viewed on a white light plate and a transilluminator to photograph the protein profiles. The gel is processed for Western blot transfer of proteins onto a nitrocellulose membrane, and upon completion, the protein profiles on the membrane are viewed and photographed as stated above. The membrane can then be processed for immunostaining as per the standard procedure. Thus, the staining procedure using Alta is simple, rapid (without any need of destaining) and cost-effective.
SDS–polyacrylamide gels Nitrocellulose membrane Western blotting Alta Protein stain
This is a preview of subscription content, log in to check access.
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
This work is supported by a research grant from the Department of Atomic Energy, Govt. of India, Bhabha Atomic Research Centre, Mumbai (BRNS Grant number 2007/37166/BRNS) and a grant from the Department of Science and Technology (DST), Govt. of India under the DST PURSE Programme. We wish to thank Dr. Jayant Londhe for his assistance in initiating some preliminary experiments.
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefGoogle Scholar
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76:4350–4354CrossRefPubMedPubMedCentralGoogle Scholar
Pal JK, Sarkar A, Katoch B (2001) Detergent-mediated destaining of Coomassie brilliant blue-stained SDS polyacrylamide gels. Indian J Exp Biol 39:95–97PubMedGoogle Scholar
Pal JK, Godbole D, Sharma K (2004) Staining of proteins on SDS polyacrylamide gels and on nitrocellulose membranes by alta, a colour used as a cosmetic. J Biochem Biophys Methods 61:339–347CrossRefPubMedGoogle Scholar
Kurien BT, Scofield RH (1998) Heat mediated quick Coomassie blue protein staining and destaining of SDS PAGE gels. Indian J Biochem Biophys 35:385–389PubMedGoogle Scholar
Merril CR, Goldman D, Sedman SA et al (1981) Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science 211:1437–1438CrossRefPubMedGoogle Scholar
Zehr BD, Savin TJ, Hall RE (1989) A one-step low background Coomassie staining procedure for polyacrylamide gels. Anal Biochem 182:157–159CrossRefPubMedGoogle Scholar
Wu W, Welsh MJ (1996) Rapid Coomassie blue staining and destaining of polyacrylamide gels. BioTechniques 20:386–388CrossRefPubMedGoogle Scholar
Wu M, Stockley PG, Martin WJ II (2002) An improved Western blotting technique effectively reduces background. Electrophoresis 23:2373–2376CrossRefPubMedGoogle Scholar
Antharavally BS, Carter B, Bell PA et al (2004) A high-affinity reversible protein stain for western blots. Anal Biochem 329:276–280CrossRefPubMedGoogle Scholar