Root D D, Wang K
Department of Chemistry and Biochemistry, Clayton Foundation Biochemical Institute, University of Texas, Austin 78712.
Anal Biochem. 1993 Feb 15;209(1):15-9. doi: 10.1006/abio.1993.1077.
Copper staining of protein blots has previously been shown to detect proteins rapidly, quantitatively, and sensitively. The sensitivity of this assay can now be enhanced nearly 10-fold with an additional step of silver staining. The silver-enhanced assay retains many of the virtues of copper staining, including a quantitative optical response with the amount of adsorbed protein, low variation between proteins, and reversibility. In addition, both copper staining and silver-enhanced copper staining do not detect nucleic acid, thus allowing for selective measurement of nucleic acid binding proteins or protein contamination in nucleic acid preparations. Silver-enhancement requires only a 5-min incubation with a single reagent. Since copper staining is performed first, it can be used to determine if the greater sensitivity provided by silver enhancement is desirable. For quantitation, an inexpensive reflectance scanning densitometer utilizing optical scanners and NIH Image is described.
蛋白质印迹的铜染色此前已被证明能够快速、定量且灵敏地检测蛋白质。现在，通过额外的银染步骤，该检测方法的灵敏度可提高近10倍。银增强检测法保留了铜染色的许多优点，包括与吸附蛋白量呈定量光学响应、蛋白质间差异小以及具有可逆性。此外，铜染色和银增强铜染色均无法检测核酸，因此可用于选择性测量核酸结合蛋白或核酸制剂中的蛋白质污染。银增强仅需与单一试剂孵育5分钟。由于首先进行铜染色，因此可用于确定银增强所提供的更高灵敏度是否可取。对于定量分析，本文描述了一种利用光学扫描仪和NIH Image的廉价反射扫描密度计。
