AN 8-FOLD TO 10-FOLD ENHANCEMENT IN SENSITIVITY FOR QUANTITATION OF PROTEINS BY MODIFIED APPLICATION OF COLLOIDAL GOLD
AN 8-FOLD TO 10-FOLD ENHANCEMENT IN SENSITIVITY FOR QUANTITATION OF PROTEINS BY MODIFIED APPLICATION OF COLLOIDAL GOLD
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DOI：
10.1016/0003-2697(88)90319-3
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发表时间：
1988-02-01
影响因子：
2.9
通讯作者：
GABIUS, HJ
中科院分区：
文献类型：
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作者：
CIESIOLKA, T;GABIUS, HJ
我们对C.M.Stoscheck(1987，肛门)的高灵敏蛋白质分析方法进行了改进。生物化学。160、301-305)，导致灵敏度进一步提高8到10倍。该分析响应于蛋白质数量，检测下限为1 ng，只需将现在组织化学和蛋白质印迹中常用的胶体金溶液添加到蛋白质样品中，然后在几分钟内简单地测量590 nm处的吸光度变化。通过增加胶体金的浓度，使用用0.01%聚乙二醇稳定并调节到pH 3.8的金溶胶，并将该分析方法调整到微滴定板上，这种类型的分析可以用于可靠地测定完整纳克范围内的蛋白质。因此，这种分析方法在快速、灵敏、昂贵和不受许多实验室试剂干扰等方面优于其他分析方法，尽管像其他方法一样，它也存在着不同蛋白质反应差异的缺点，这是染料结合分析所固有的。
We have modified the highly sensitive protein assay of C. M. Stoscheck (1987, Anal. Biochem. 160, 301-305), resulting in a further 8- to 10-fold enhancement of sensitivity. This assay, responding to protein quantities with a detection limit of 1 ng, involves the single step of addition of colloidal gold solution, as now commonly used in histochemistry and protein blotting, to the protein sample, followed by simple measurement of the change in absorbance at 590 nm within minutes. By increasing the concentration of the colloidal gold, by using gold sol that has been stabilized with 0.01% polyethylene glycol and adjusted to pH 3.8, and by adapting the assay to microtiter plates, this type of assay can be applied to reliably determine proteins in the complete nanogram range. This assay therefore compares favorably to other assay procedures interms of rapidity, sensitivity, expense, and lack of interference by many laboratory reagents, although like the others it suffers from the drawback of differences in response of different proteins, which is inherent in dye-binding assays.