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本文介绍用3%～25%梯度 SDS—聚丙烯酰胺凝胶电泳(3%～25%Gradient SDS-PAGE)测定人类胚胎晶体蛋白的方法,对照试验了蛋白质样品在电泳前的不同预处理条件和预处理时间,发现当水溶性晶体蛋白在电泳前与样品处理液混合后,在37℃保温3小时,在分子量为45KDa 附近有2—3条亚基谱带的颜色明显加深,而在100℃沸水中保温2分钟,这些谱带颜色变浅,在29KDa 附近的谱带颜色加深。从这些现象我们初步推断出在胚胎晶体水溶性蛋白中存在一种酶,这种酶在一定程度上能抑制SDS 和巯基乙醇对某些晶体蛋白质肽链的降解作用。但在晶体水不溶性蛋白中,则不存在这一种酶。本文还介绍了使用硼酸——NaOH(0.1%SDS)电极缓冲液代替 Gly-Tris(0.1%SDS)电极缓冲液进行 SDS-PAGE 测定的试验。
This article describes a method for the determination of human embryonic lens proteins using a 3% to 25% gradient SDS-polyacrylamide gel electrophoresis (3% to 25% Gradient SDS-PAGE), comparing different pretreatment conditions for protein samples before electrophoresis and Pretreatment time and found that when the water-soluble crystal protein electrophoresis before mixing with the sample processing solution, incubated at 37 ℃ for 3 hours, the molecular weight of 45KDa in the vicinity of 2-3 subunit bands significantly deepened the color at 100 ℃ After incubation in boiling water for 2 minutes, these bands became lighter in color and deepened in the vicinity of 29 kDa. From these phenomena, we initially concluded that there is an enzyme in the crystal protein of embryonic water-soluble protein, which can inhibit the degradation of some crystal protein peptides chains to a certain extent by SDS and mercaptoethanol. However, in crystalline water-insoluble proteins, this enzyme is absent. This article also describes the test for the SDS-PAGE assay using a boric acid-NaOH (0.1% SDS) electrode buffer instead of a Gly-Tris (0.1% SDS) electrode buffer.