Glyoxalase I (GLO I) Phenotyping by Isoelectric Focusing

Abstract

The polymorphism of the red cell enzyme glyoxalase I (GLO-I), first described by Kömpf et al. [1] using starch gel electrophoresis, was investigated by isoelectric focusing (IEF) on polyacrylamide plates at pH 4-5 (LKB, PAG plates). Briefly, 20 μl red cell lysate was applied near the cathode. Following the focusing, the gels were incubated for 20 minutes at 37 °C in a 0.2 M phosphate buffer containing 257 mM methylglyoxal and 16.3 mM reduced glutathione. The detection gel consisted of 1% agarose in 0.1 M Tris buffer with 0.06 mM DCIP and 2.4 mM MTT poured on a GelBond film.