Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250.
An improved procedure for staining of proteins following separation in polyacrylamide gels is described which utilizes the colloidal properties of Coomassie Brilliant Blue G-250 and R-250. The new method is based on addition of 20% v/v methanol and higher concentrations of ammonium sulfate to the staining solution previously described. The method combines the advantage of much shorter staining time with high sensitivity, a clear background not requiring destaining, stepwise staining, and stable fixation after staining. The method has been applied to staining of polyacrylamide gels after sodium dodecyl sulfate-electrophoresis and isoelectric focusing in carrier ampholyte-generated pH gradients.
Neuhoff
V
V
Max-Planck-Institut für experimentelle Medizin, Göttingen, Federal Republic of Germany.
Arold
N
N
Taube
D
D
Ehrhardt
W
W
D000180:Acrylic Resins; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D005782:Gels; D007525:Isoelectric Focusing; D008027:Light; D011506:Proteins / Q000032:analysis*; D012394:Rosaniline Dyes; D012542:Scattering, Radiation; D013194:Staining and Labeling
D000180:Acrylic Resins; D005782:Gels; D011506:Proteins; D012394:Rosaniline Dyes; C016680:polyacrylamide gels; C024757:Coomassie brilliant blue R; C004692:coomassie Brilliant Blue