Sears D A, Lewis P C
J Lab Clin Med. 1980 Aug;96(2):318-27.
A method utilizing sodium dodecyl sulfate--polyacrylamide gel electrophoresis for the sepration of the alpha and beta chains of hemoglobin has been adapted to quantify hemoglobin chains by simultaneous electrophoresis of globin standards with unknowns, staining with Coomassie blue, densitomeric scanning, and planimetry. This method has been used to quantify the globin chains bound to the human red cell membrane during sterile incubation and ATP depletion in vitro. In thoroughly washed (6-step) ghosts of incubated cells; (1) more beta than alpha chains were bound (beta/alpha ratio 1.28); (2) only about one third of the bound chains contained heme; and (3) globin accounted for less than 20% of the "excess" protein present in the incubated cells compared to fresh cells. Four-step ghosts contained more hemoglobin, a smaller proportion of heme-free alpha and beta chains, and approximately equal numbers of alpha and beta chains (beta/alpha ratio 1.09).
一种利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离血红蛋白α链和β链的方法，已通过将珠蛋白标准品与未知样品同时进行电泳、用考马斯亮蓝染色、光密度扫描和平面测量来定量血红蛋白链。该方法已用于在体外无菌孵育和ATP耗竭期间定量与人红细胞膜结合的珠蛋白链。在经过充分洗涤（6步）的孵育细胞血影中：（1）结合的β链比α链多（β/α比率为1.28）；（2）仅约三分之一的结合链含有血红素；（3）与新鲜细胞相比，珠蛋白占孵育细胞中“过量”蛋白质的比例不到20%。四步血影含有更多的血红蛋白，无血红素的α链和β链比例更小，且α链和β链数量大致相等（β/α比率为1.09）。
