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简化的不连续梯度聚丙烯酰胺凝胶电泳系统

System for simplified discontinuous-gradient polyacrylamide-gel electrophoresis.

作者信息

Zais D P, Roberts R C

出版信息

Clin Chem. 1977 Mar;23(3):590-2.

PMID:65234
Abstract

We describe a modified polyacrylamide gel electrophoresis method with which plasma proteins are excellently resolved in 4 h. We made a three-layer discontinuous gradient gel with concentrations of 80,60, and 45 g of polyacrylamide per liter of tris(hydroxymethyl)aminomethane/ethylenediaminetetraacetic acid/borate buffer, pH 9.2. Time and material needed in the preparation of the gel was greatly decreased, a well-resolved (22-25 bands) pattern was obtained in 1 h, and the gel slabs were easier to remove from the cell than were gels prepared according to the manufacturer's specifications. We saw a marked shift in the higher polymers of haptoglobin type 2-2 toward the cathode in this system, and also a clearly resolved double band of alpha2-macroglobulin in whichthe more anodal band appears to represent alpha2-macroglobulin/protease complex.

摘要

我们描述了一种改良的聚丙烯酰胺凝胶电泳方法,利用该方法可在4小时内出色地分离血浆蛋白。我们制备了一种三层不连续梯度凝胶,其在每升pH 9.2的三(羟甲基)氨基甲烷/乙二胺四乙酸/硼酸盐缓冲液中的聚丙烯酰胺浓度分别为80、60和45克。制备凝胶所需的时间和材料大幅减少,1小时内即可获得条带清晰的图谱(22 - 25条带),并且与按照制造商规格制备的凝胶相比,该凝胶板更易于从电泳槽中取出。在该系统中,我们观察到触珠蛋白2 - 2型的高分子聚合物明显向阴极迁移,同时还清晰地分辨出α2 - 巨球蛋白的双条带,其中阳极条带似乎代表α2 - 巨球蛋白/蛋白酶复合物。

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