Abstract
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.
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Acknowledgment
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.
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Pal, J.K., Berwal, S.K., Soni, R.N. (2018). A Single-Step Simultaneous Protein Staining Procedure for Polyacrylamide Gels and Nitrocellulose Membranes by Alta During Western Blot Analysis. In: Kurien, B., Scofield, R. (eds) Protein Gel Detection and Imaging. Methods in Molecular Biology, vol 1853. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8745-0_12
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DOI: https://doi.org/10.1007/978-1-4939-8745-0_12
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