pH Gradient flattening in isoelectric focusing in long polyacrylamide gels

Abstract
Isoelectric focusing in 60 cm long gels improves resolution in multiple‐component systems by providing very shallow, linear pH gradients. Broad range ampholytes available commercially or synthesized in our laboratory develop pH gradients of 0.1 pH/cm at voltages of 1500 to 4500 V, with running times of 25–30 h. Shallower gradients result with narrow range ampholytes or separator isoelectric focusing. Hemoglobin, human serum albumin, and an α‐N‐acetylgalactosaminidase have been used for defining this system.