QUANTITATION OF HUMAN GLOBIN CHAIN SYNTHESIS BY CELLULOSE-ACETATE ELECTROPHORESIS

被引:8
作者
SALMON, JE [1 ]
NUDEL, U [1 ]
SCHILIRO, G [1 ]
NATTA, CL [1 ]
BANK, A [1 ]
机构
[1] COLUMBIA UNIV, DEPT HUMAN GENET & DEV, NEW YORK, NY 10032 USA
关键词
D O I
10.1016/0003-2697(78)90825-4
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We have developed a method for the separation and quantitation of human α-, β-, and γ-globins utilizing cellulose acetate electrophoresis. The relative rates of synthesis of globin chains in reticulocytes in peripheral blood is determined by: (i) incubating intact cells with [35S]methionine; (ii) preparing globin from the hemolysates; (iii) performing electrophoresis of the globin on cellulose acetate strips; and (iv) autoradiography or direct determination of the radioactivity incorporated into each globin chain. The method is simple and rapid, requires only small amounts of hemolysate (30 μg of globin), and provides excellent resolution and reproducible quantitation of α-, βA-, βS-, and γ-globin chains for up to 24 peripheral blood samples at one time. Measurements by this method in patients with thalassemia variants and sickle-cell disorders correlate well with analysis of the same samples by carboxymethyl cellulose chromatography. This methodology may permit more widespread analysis of globin synthesis in the thalassemia syndromes and may also be useful in the analysis of globins synthesized from human globin mRNA in cell-free systems. © 1978.
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收藏
页码:146 / 157
页数:12
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