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Tritium/PPO gel fluorographic efficiency is reduced by Coomassie Blue staining
Author(s) -
Higgins Ratchford C.,
Dahmus Michael E.
Publication year - 1982
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150030407
Subject(s) - coomassie brilliant blue , chemistry , chromatography , staining , stain , polyacrylamide gel electrophoresis , gel electrophoresis , silver stain , methanol , electrophoresis , biochemistry , enzyme , microbiology and biotechnology , organic chemistry , biology , genetics
When [ 3 H]‐labeled proteins are analyzed by polyacrylamide gel electrophoresis, the sensitivity of fluorograhic detection following gel impregnation with PPO (2,5‐diphenyloxazole) in DMSO (dimethylsulfoxide) is significantly reduced if the gel has been stained with Coomassie Brilliant Blue R‐250. Since the reduction in fluorographic efficiency depends on the concentration of stain, which in turn depends upon protein concentration, substantial errors may enter into quantitative measurements of fluorograms depending on sample composition. Another reduction in fluorographic efficiency results from reduced penetrability of the gels to PPO when a build‐up of methanol in DMSO and DMSO/PPO solutions is allowed. This loss of fluorographic sensitivity should be sample independent, however.