Abstract
The zone of stacked protein in polyacrylamide gel electrophoresis (PAGE) generated by a multiphasic (discontinuous) buffer system was analyzed by continuous optical scanning during electrophoresis. In agreement with theoretical prediction, zone width was found to be constant with time; band area was linearly related to protein load; band dispersion, measured as the standard deviation (σ) of the concentration profile, was independent of current density between 1 and 8 mA/cm 2; and σ decreased with increasing ionic strength of the gel buffer. The protein concentration within the stack ranged from 13 to 34 mg/ml for β-lactoglobulin B of 36,000 mw. In lieu of the predicted uniform distribution (“square-wave” function) for peak shape, the observed values of kurtosis and skewness indicated a slightly flattened Gaussian profile of the protein concentration in the stack.
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