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Analytical micro-preparative electrophoresis: quantitation of phosphoglucose isomerase isoenzymes
1Charles Salt Research Centre, Robert Jones and Agnes Hunt Orthopaedic Hospital, Oswestry, Salop, England.
Abstract:
A simple micro-preparative electrophoresis apparatus is described using a thin vertical slab gel and continuous elution through a narrow horizontal slot. Phosphoglucose isomerase (PGI) isoenzymes in lysates of 0.2 to 100 microliters of erythrocytes were separated using this system and their activities determined using an on-line, air-segmented, continuous flow assay. Ninety-five percent recovery of the separated isoenzymes after electrophoresis was achieved and the factors influencing recovery of PGI and hemoglobin were investigated. Thus, quantitative recovery has been achieved without any loss of resolution. Up to 29 successive separations of PGI isoenzymes have been carried out on a single gel.