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Characterisation of the S-Carboxymethyl Extract of Wool Using Small Format Alkaline Urea-PAGE/SDS-PAGE Followed by Silver Staining
Abstract:Abstract

Alkaline urea-PAGE/SDS-PAGE, when used in a novel format which was significantly smaller than that employed by earlier workers, and when followed by silver staining, resulted in the detection of excellently resolved protein components from S-carboxymethyl reductive extracts of very small wool samples, even samples as small as an individual wool fibre. The silver stained two-dimensional gel patterns exhibited significant improvements compared to the fluorograph gel patterns of earlier workers based on the presence of radiolabel incorporated within the S-carboxymethyl moiety. The silver staining resulted in the visualisation of numerous protein components, and in the region of the gel pattern expected to contain the high-sulfur protein fraction, there appeared more major components than the number of high-sulfur protein components usually displayed in fluorograph patterns. The relative amount of the high-sulfur protein fraction in the final silver stained gel pattern could be boosted, if desired, if the gel was loaded not with the whole wool extract but with the filtrate resulting from the passage of the whole extract through a centrifugal ultrafilter.
Keywords:S-Carboxymethyl wool proteins  two-dimensional gas electrophoresis  alkaline urea-PAGE/SDS-PAGE  small format/silver staining  improved resolution  interpretation of gel pattern
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