Specific indication of hemoproteins in polyacrylamide gels using a double-staining process.
Hemoproteins were revealed in polyacrylamide gels in the presence of sodium dodecyl sulfate by staining with different benzidine derivatives. When the protein samples were treated with either beta-mercaptoethanol or dithiothreitol, a significant decrease in peroxidase activity of the proteins possessing noncovalently bound heme led to diminished staining. However, when Coomassie blue R-250 staining followed the hemespecific stain it was observed that the hemoprotein bands stained more intensely than duplicate sample bands that had been stained only with the Coomassie blue R-250. This staining property allows the indication of hemoproteins in gels even after the peroxidase yield has been significantly depleted by reducing agents.
D001560:Benzidines; D003574:Cytochrome c Group / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006418:Heme / Q000032:analysis; D006420:Hemeproteins / Q000032:analysis*; D006454:Hemoglobins / Q000032:analysis; D007527:Isoenzymes; D009195:Peroxidase; D010544:Peroxidases; D012394:Rosaniline Dyes; D012967:Sodium Dodecyl Sulfate; D013194:Staining and Labeling
D001560:Benzidines; D003574:Cytochrome c Group; D006420:Hemeproteins; D006454:Hemoglobins; D007527:Isoenzymes; D012394:Rosaniline Dyes; D012967:Sodium Dodecyl Sulfate; D006418:Heme; D010544:Peroxidases; D009195:Peroxidase; C004692:coomassie Brilliant Blue