超氧化物歧化酶和乳酸脱氢酶在凝胶板上同时显色的方法

本文报道一种能使乳酸脱氢酶(LDH)和超氧化物歧化酶(SOD)同工酶在同一块电泳凝胶板上经同一种染色液一次处理达到同时显色的方法。显色结果,LDH为有色区带,SOD为无色透明区带,两者一目了然。而且前者泳动慢,后者泳动快,两种区带互不重迭,不发生干扰,结果可靠。此法不需光照,显色效率高,有利于同一样品同时进行此两种同工酶的电泳分析,达到一举两得的效果。

Simultaneous Development of the Zymograms for Superoxide Dismutase and Lactate Dehydrogenase on a Single Gel Slab

This paper reports a method of simultaneous development of the zymograms for superoxide dismutase (SOD) and lactate dehydrogenase (LDH) isozymes on the same gel slab by means of staining once in a single staining solution after vertical slab electrophoresis of tissue homogenates. The two zymograms can be distinguished by positive staining reaction (for LDH) and negative staining reaction (for SOD), according to the appearance of colored zone (LDH) and colorless or transparent zone (SOD) on a stained background. They can also be exactly identified by their mobilities, because the migratory speed of SOD zones toward positive electrode are faster than that of LDH ones. Thus, the two zymograms are not likely to overlep. Test of cyanide inhibition show that the SOD zones mentioned above belong to Cu/Zn-SOD. To find out the effect of each reagent on simultaneous development, the staining efficiency when a particular reagent is absent is compared with that when it is present. The paper discussed the source of the superoxide anion radicals in the staining system. It also discussed the two different kinds of mechanism of the nitro blue tetrazolium being reduced to a blue formazan, i.e., by the superoxide anion radicals in regions where SOD is not present, and by the hydrogen ion in regions where LDH is present.