Vincent R, Nadeau D
Anal Biochem. 1983 Dec;135(2):355-62. doi: 10.1016/0003-2697(83)90696-6.
A simple procedure for the determination of cellular proteins in Percoll-containing samples is described. Percoll precipitated when particulate proteins were solubilized by dilution of the samples in a NaOH-Triton X-100 mixture. After centrifugation at high speed (12,000g), the supernatant was assayed for proteins with the Coomassie brilliant blue dye-binding assay. With an automatic spectrophotometer, 50-microliter aliquots gave a linear response between 0 and 3 micrograms of bovine serum albumin. After a fivefold dilution in the alkali-detergent mixture, proteins in samples containing up to at least 60% Percoll can be accurately quantitated on a standard curve prepared in the absence of Percoll. Because the sensitivity of the assay was better than 100 ng, the procedure outlined in this paper can also be used as a general protein micromethod.
本文描述了一种用于测定含Percoll样品中细胞蛋白质的简单方法。当通过在NaOH - Triton X - 100混合物中稀释样品使颗粒状蛋白质溶解时，Percoll会沉淀。高速离心（12,000g）后，用上清液通过考马斯亮蓝染料结合法测定蛋白质。使用自动分光光度计，50微升等分试样在0至3微克牛血清白蛋白之间呈现线性响应。在碱洗涤剂混合物中进行五倍稀释后，含有至少60% Percoll的样品中的蛋白质可以在不含Percoll的情况下制备的标准曲线上进行准确定量。由于该测定法的灵敏度优于100纳克，本文所述方法也可作为一种通用的蛋白质微量测定法。
