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Alternative methods for fixing and staining gliadins in polyacrylamide gels
Author(s) -
Clements Robert L.
Publication year - 1990
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.1150110204
Subject(s) - staining , polyacrylamide , chemistry , polyacrylamide gel electrophoresis , chromatography , biophysics , biochemistry , biology , polymer chemistry , genetics , enzyme
Staining efficiencies of Coomassie Brilliant BlueG‐250 (CBB G‐250) and Coomassie Brilliant Blue R‐250 (CBB R‐250) in various media were studied in efforts to reduce or eliminate requirements for trichloroacetic acid (TCA). Stained gels were compared with gels stained with CBB R‐250 in 12% TCA and evaluated for overall stain and background. Because of qualitative effects, stain intensities of low‐ and highmobility gliadins were also evaluated. Results indicated gliadins are fixed under a wide range of conditions, permitting adjustment of conditions to provide optimum staining. CBB G‐250 and R‐250 in tap water fixed and stained most gliadins. Best results were obtained with CBB G‐250 in 2% TCA, in 2% TCA containing 5% sodium sulfate, and in 2% and 5% phosphoric acid containing 5% sodium chloride or 5% sodium sulfate. Gels stained in these media were more easily observed during staining and more easily destained than gels stained in CBB R‐250 in 12% TCA.