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本文报告一种人红细胞膜血型糖蛋白A(Glycophorin A)的分离纯化方法。血库血(AB型),采用0.9％NaCl及旋离手段去除白细胞、血小板和血浆蛋白等,洗净的红细胞用蒸馏水溶破,离心沉淀得人红细胞膜制剂。膜制剂用热酚抽提,得粗糖蛋白。再用DEAE—Sephadex A-25和SephadexG-100柱层析可从粗糖蛋白中获得纯血型糖蛋白A。 纯化的血型糖蛋白A经SDS—PAG(十二烷基硫酸钠-聚丙烯酰胺凝胶)电泳鉴定,呈现一条蛋白带,PAS(过碘酸—Schiff氏试剂)染色为阳性,与全膜蛋白SDS—PAG电泳相比相当于PAS—1的位置。用SDS-PAG电泳测得分子量约为60000。
This article reports a method for the isolation and purification of human erythrocyte glycoprotein A (Glycophorin A). Serum blood (AB type), using 0.9% NaCl and spin-off means to remove leukocytes, platelets and plasma proteins, washed red blood cells dissolved in distilled water, centrifuged to obtain human erythrocyte membrane preparation. Membrane preparation with hot phenol extraction, was crude glycoprotein. Pure glycoprotein A was then obtained from the crude glycoprotein using DEAE-Sephadex A-25 and Sephadex G-100 column chromatography. Purified glycophorin A was identified by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel) electrophoresis, showing a protein band, PAS (periodic acid-Schiff’s reagent) staining was positive, and the whole membrane protein SDS-PAG electrophoresis compared to the position corresponding to PAS-1. SDS-PAG electrophoresis measured molecular weight of about 60,000.