551 Biji T. Kurien and R. Hal Scofield (eds.), Protein Electrophoresis: Methods and Protocols, Methods in Molecular Biology, vol. 869, DOI 10.1007/978-1-61779-821-4_49, © Springer Science+Business Media, LLC 2012 Chapter 49 A Single-Step Simultaneous Protein Staining Procedure for Polyacrylamide Gels and Nitrocellulose Membranes by Alta During Western Blot Analysis Jayanta K. Pal, Sunil K. Berwal, and Rupali N. Soni Summary 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 and 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. Key words: SDS polyacrylamide gels, Nitrocellulose membrane, Western blotting, Alta, Protein stain Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (1) and western blot analysis (2) have become a part and parcel of research in life sciences. Considering this reality, there has been a continuous effort towards developing better versions of these pro- cedures. Among others, detection of proteins in SDS polyacrylam- ide gels and also on nitrocellulose membranes, both of which are important in the western blot analysis, has attracted much attention of researchers involved in improvement of these techniques (3–5). Along this line, a number of protein staining procedures are avail- able in the literature for staining proteins independently on poly- acrylamide gels and on nitrocellulose/PVDF membranes (6–11). The commonly used stains for gels and membranes are Coomassie Brilliant Blue (R-/G-250) and Ponceau Red S, respectively. 1. Introduction