The localization of spectrin on the inner surface of human red blood cell membranes by ferritin-conjugated antibodies.
The localization of spectrin on the inner surface of human red blood cell membranes by ferritin-conjugated antibodies.
复制标题
DOI：
10.1083/jcb.51.1.265
复制
发表时间：
1971-10
期刊：
影响因子：
--
通讯作者：
Singer SJ
中科院分区：
文献类型：
--
作者：
Nicolson GL;Marchesi VT;Singer SJ
Spectrin是哺乳动物红细胞膜制备物的主要蛋白质成分，已通过利用特异性铁蛋白缀合抗体的技术定位于人红细胞膜的内表面，并在溶血后不久固定膜，以允许铁蛋白抗体标记物穿透。血影蛋白的标记通过以下标准显示为特异性的。(a)非同源铁蛋白偶联抗体不特异性结合到任一膜表面。(b)用过量的未缀合的抗血影蛋白抗体阻断膜结合血影蛋白阻止铁蛋白抗体标记。(c)通过用含有乙二胺四乙酸盐和β-巯基乙醇的低离子强度缓冲液处理膜制备物来去除血影蛋白，防止了特异性铁蛋白缀合抗体的标记。
Spectrin, a major protein constituent of mammalian red blood cell membrane preparations, has been localized on the inner surface of human red blood cell membranes by techniques that utilized specific ferritin-conjugated antibodies and fixation of membranes shortly after hemolysis so as to allow penetration of the ferritin-antibody labels. The labeling of spectrin was shown to be specific by the following criteria. (a) Nonhomologous ferritin-conjugated antibodies did not specifically bind to either membrane surface. (b) Blocking the membrane-bound spectrin with excess unconjugated antispectrin antibodies prevented ferritin-antibody labeling. (c) Removal of spectrin by treating the membrane preparation with a low ionic strength buffer containing ethylenediaminetetraacetate and β-mercaptoethanol prevented labeling by specific ferritin-conjugated antibodies.