Incorporation of methylamine in an ultrasensitive silver stain for detecting protein in thick polyacrylamide gels

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Abstract
An ultrasensitive silver stain is described for the detection of proteins in 4–20 % w/v polyacrylamide gradient gels of 3mm thickness, Maximal sensitivity was achieved by substituting methylamine for ammonia in the diamine preparation and adopting a strategy of overstain followed by destain. The prediamine steps were performed at 60°C to conserve time, and the intensity of final background stain was reduced by altering the method of diamine preparation. From our experience, the technique described is more sensitive and reliable than previously published methods when applied to gels of this thickness, In any case, the sensitivity in our hands was at least equal to that of the newly described photochemical technique [7].