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Analysis of differences between coomassie blue stain and silver stain procedures in polyacrylamide gels: conditions for the detection of calmodulin and troponin C
Authors:M Schleicher  D M Watterson
Institution:1. Laboratory of Cellular and Molecular Physiology, Howard Hughes Medical Institute, Vanderbilt University, Nashville, Tennessee 37232 USA;2. Department of Pharmacology, Vanderbilt University, Nashville, Tennessee 37232 USA
Abstract:It is reported that the conditions used in some silver stain procedures can fail to detect calmodulin, troponin C, and other proteins with similar physical properties. Conditions are described that allow the reproducible detection of these proteins. Two phenomena are described: (1) lack of protein staining when treatment with glutaraldehyde is omitted from the protocol, and (2) loss of small proteins from the gel matrix during prolonged washing procedures. These data directly demonstrate that the use of some silver staining protocols can result in misleading data in biological studies and provide an explanation for at least one class of proteins of how silver staining and Coomassie blue staining of gels can give different results.
Keywords:calmodulin  troponin C  electrophoresis  Coomassie blue stain  silver stain  calcium-binding proteins
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