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Silver stain for detecting 10-femtogram quantities of protein after polyacrylamide gel electrophoresis
Authors:K Ohsawa  N Ebata
Affiliation:1. Pure and Applied Biochemistry, Chemical Center, University of Lund, P.O. Box 740, S-220 07 Lund, Sweden;2. Department of Biochemistry, Trinity College, Dublin University, Dublin, Ireland
Abstract:A rapid and highly sensitive silver stain and color stain were developed for visualizing proteins. The procedure is simple and the bands were clear. This silver stain detects 100 pg quantities of proteins. In order to stain quickly, sensitively, and sharply a protein matrix in a gel, the repeated shrinkage and swelling gel was developed with a hyper- and hypotonic solution to remove the sodium dodecyl sulfate (SDS) from SDS-protein complex and to generate influx of staining solution into the gel. We have found that the silver staining method with the repeated exposure to hyper- and hypotonic solution and a narrow well produced 10 fg order of proteins.
Keywords:silver stain  protein polyethylene glycol  sodium dodecyl sulfate  gel shrinkage  gel swelling
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