Abstract
A method for isoelectric focusing in polyacrylamide gels (IFPA) has been described. A 3.7% T polyacrylamide gel allowed for isoelectric focusing of hemoglobin and cyanogen bromide fragments of both collagen and hemoglobin in the absence of significant molecular sieving and provided satisfactory mechanical stability. Support of the gels was enhanced by polyacrylamide coating of tubes, by use of a membrane support, and by hydrostatic equilibration. Runs for 8 hr at 200 V/6 cm at 5°C provided optimal separation. Preparative IFPA was conducted in tubes of up to 18 mm diameter with loads of up to 2 mg per band. Protein was recovered quantitatively from the gel slices.
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