Using coomassie blue to stabilize H2O2-guaiacol stained peroxidases on polyacrylamide gels.
Shimoni and Reuveni [1] have reported a procedure which uses Coomassie Brilliant Blue R-250 staining and destaining to stabilize H2O2-guaiacol stained peroxidases on polyacrylamide gels and at the same time detects nonspecific protein bands. The procedure has the advantage that the otherwise orange and unstable peroxidase bands are visualized as blue bands and it, therefore, facilitates laser densitometry of electrophoretically separated peroxidases. In the modification of the procedure reported here, the Coomassie staining reagent was adjusted and the destaining step was eliminated. The modified procedure has added advantages: it simplifies the procedure, increases the consistency of the results across gels, prevents or reduces the staining of nonspecific proteins, and still gives excellent resolution of stable bands which can be used to quantify peroxidase activity.
Fieldes
M A
MA
Department of Biology, Wilfrid Laurier University, Waterloo, Ontario, Canada.
D004591:Electrophoresis, Polyacrylamide Gel; D006139:Guaiacol; D006861:Hydrogen Peroxide; D007527:Isoenzymes / Q000032:analysis; D010544:Peroxidases / Q000032:analysis*; D012394:Rosaniline Dyes; D013194:Staining and Labeling
D007527:Isoenzymes; D012394:Rosaniline Dyes; C024757:Coomassie brilliant blue R; D006139:Guaiacol; D006861:Hydrogen Peroxide; D010544:Peroxidases