人红细胞NADH-细胞色素b5还原酶是使高铁血红蛋白还原的主要酶类, 其缺陷将导致遗传性高铁血红蛋白血症. 目前, 主要通过分光光度法测定b5还原酶活性. 我们将b5还原酶抗体点于硝酸纤维膜上, 以此捕获并富集红细胞胞浆b5还原酶. 有b5还原酶活性的斑点用噻唑蓝染色. 此法简单直观, 可用于b5还原酶的定性和半定量测定, 为遗传性高铁血红蛋白血症的诊断提供了一种新的实验手段.
Human erythrocyte NADH-cytochrome b5 reductase, or b5 reductase, plays a major role in the reduction of methemoglobin,deficiency of which will lead to hereditary methemoglobinemia. Deterrnination of b5 reductase activity is usually done by spectrophotometry. A new method has been developed for the qualitative and semi-quantitative detection of b5 reductase activity, in which antibodies against b5 reductase was dot-blotted onto nitrocellulosemembrane, and this in turn was used to capture and enrich b5 reductase activity were visualized with the precipitable substrate MTT, or 3-(4, 5-dimethyl thiazolyl-2)-2, 5-diphenyl-tetrozolium bromide. Being straightforward and easy to follow, this method provides a new approach for the diagnosis of hereditary methemoglobinemia.
兰风华,唐玉钗,朱忠勇,吴玉水.膜上斑点洁显示人红细胞b5还原酶活性[J].生物化学与生物物理进展,1996,23(4):379-380
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