'Catalysts' for polyacrylamide gel polymerization and detection of proteins by silver staining.
The crosslinker diacrylyl-piperazine produces polyacrylamide gels which display improved electrophoretic separation of proteins and better physical strength. It also produces gels with improved detection of proteins by ammoniacal silver staining by reducing the background. This reduced background provided us with an opportunity to investigate residual background staining caused by the catalytic reagents utilized in the polymerization of acrylamide gels. The commonly used catalyst system, tetramethyl-ethylenediamine and ammonium persulfate was shown to be responsible for the yellow staining background found after a prolonged development time with silver staining. An alternate catalyst system has been designed to decrease further the formation of this background staining. Dimethyl-piperazine or tetramethylethylenediamine, potassium or ammonium persulfate, and sodium thiosulfate are shown to provide for gels which have excellent mechanical and staining characteristics. These catalytic systems produce little background staining despite prolonged development time with the ammoniacal silver stain, and they reduce background staining with the dichromate silver stain.
Hochstrasser
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DF
Section of Biochemical Genetics, National Institute of Mental Health, Bethesda, Maryland 20892.
Merril
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CR
D000180:Acrylic Resins / Q000138:chemical synthesis*; D002384:Catalysis; D004591:Electrophoresis, Polyacrylamide Gel; D010084:Oxidation-Reduction; D011506:Proteins / Q000032:analysis*; D012834:Silver; D013194:Staining and Labeling
D000180:Acrylic Resins; D011506:Proteins; C000212:ammoniacal silver; C016680:polyacrylamide gels; D012834:Silver