Kinetic silver staining and quantification of proteins adsorbed to microtiter plates.
Kinetic silver staining and quantification of proteins adsorbed to microtiter plates.
复制标题
吸附到微量滴定板的蛋白质的动力学银染色和定量。
DOI：
10.1006/abio.1993.1134
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发表时间：
1993
影响因子：
2.9
通讯作者：
Wang,K
中科院分区：
文献类型：
--
作者：
Root,DD;Wang,K
使用银染剂检测和定量吸附到微量滴定板威尔斯孔中的蛋白质。用自动微量滴定板读数器分析银染发展的动力学。发现染色发展的滞后时间是吸附到微量滴定板孔的蛋白质量的一致指示剂。未预吸附到微量滴定板上的蛋白质不能被银有效染色。通过在非常稀的缓冲液中干燥少量蛋白质，可以完全吸附微量滴定板上的蛋白质。对于所检查的蛋白质组，不同蛋白质的灵敏度变化小于30%。动态银染法测定结果与微量滴定板吸附牛白蛋白铜染色法测定结果一致。动态银染法的精密度在40 ~ 200 ng/孔范围内最佳。在此范围内，标准偏差平均小于5%。甚至可以检测到更少量的蛋白质，并将其内推至每孔约10 ng。动态银染色方法可用于标准微量滴定板读数仪，无需特殊过滤器，并且易于适用于自动化系统。
A silver stain was used to detect and quantitate proteins adsorbed to microtiter plate wells. The kinetics of the development of the silver stain were analyzed with an automated microtiter plate reader. The lag time for stain development was found to be a consistent indicator of the amount of protein adsorbed to a microtiter plate well. Protein which was not preadsorbed to the microtiter plate was not effectively stained by silver. Complete adsorption of protein applied to the microtiter plate was possible by drying small amounts of protein in very dilute buffers. Variations in sensitivity for different proteins were less than 30% for the panel of proteins examined. Determinations from kinetic silver staining agreed with those from copper staining for bovine albumin adsorbed to microtiter plates. The precision of kinetic silver staining assay was optimal in the range of 40 to 200 ng per microtiter plate well. In this range, the standard deviations averaged less than 5%. Even smaller amounts of protein can be detected and interpolated down to approximately 10 ng per well. The kinetic silver staining method can be used on standard microtiter plate readers without special filters and is readily adaptable to automated systems.