胎盘微绒毛膜蛋白和基底膜层蛋白联合制备

The combined method of isolating human placental microvillous membrane and basilar membrane

目的探讨胎盘微绒毛膜和基底膜联合分离及其蛋白提取方法．方法采用差速离心法和蔗糖密度梯度离心法，并对所提取的膜进行电镜观察和酶学鉴定．结果电镜观察两种膜已基本分离开．酶学鉴定：ＡＫＰ酶绒毛膜为２０．０６４±８．９９６ｕ·ｍｇ－１蛋白，基底膜为０．３２３±０．１４６ｕ·ｍｇ－１蛋白．绒毛膜ＡＫＰ较基底膜高约７０倍．基底膜含少量ＡＫＰ表示有少许绒毛膜污染，表明两种膜的提取成功．结论本方法可用于联合制备胎盘绒毛膜和基底膜．

s To study for the combined isolating method of human placental microvillous membrane and basilar membrane.Methods Human placental microvillous membrane was isolated by the differential centrifugation and the basilar membrane was obstained by the discontinuous sucrose-desity-gradient centrifugation.Two kinds of membrane were identified by the electron microscopy (EM) and the marker enzymealkaline phosphatase (ALP).Results The results showed that the most of two kinds of membrane had been seprated by EM. The ALP value of microvillous membrane was 22.06± 8.996 enzyme units. mg-1. protein and the basilar membrane was 0. 323± 0. 416enzyme units mg-1. protein. The ALP of microvillous membrane was 70 times higher than that of basilar membrane. There were a few of ALP in the basilar membrane indicated that there was a little of microvillous membrane being contaminated.Conclutios It is demonstrated that the two kinds of membrane may be successfully extracted by the combined isolating method.