Lewis W S, Schuster S M
Department of Chemistry, University of Nebraska-Lincoln.
J Biochem Biophys Methods. 1990 Jul-Aug;21(2):129-44. doi: 10.1016/0165-022x(90)90060-p.
A method for the quantitative determination of immobilized proteins based on the binding and subsequent elution of Coomassie Blue R is presented. Also presented is a method for the immobilization of proteins in solution by entrapment in polyacrylamide. These entrapped proteins are then available for use in the assay method presented. Other analytical procedures can also be performed on the entrapped proteins, either alone or in combination with the protein quantitation. The dye binding and elution method presented provides a sensitive and, in most applications, rapid method for the quantitative detection of immobilized proteins. Rather than immobilization being an obstacle to the assay method, this approach utilizes the advantages of immobilization for the removal of excess reagents. Application of this approach to several types of immobilized protein are presented.
本文介绍了一种基于考马斯亮蓝R的结合及后续洗脱来定量测定固定化蛋白质的方法。还介绍了一种通过包埋于聚丙烯酰胺中来固定溶液中蛋白质的方法。这些被包埋的蛋白质随后可用于所介绍的测定方法。也可对被包埋的蛋白质单独或与蛋白质定量分析相结合进行其他分析程序。所介绍的染料结合及洗脱方法为固定化蛋白质的定量检测提供了一种灵敏且在大多数应用中快速的方法。这种方法并非将固定化视为测定方法的障碍，而是利用固定化的优势来去除过量试剂。本文展示了该方法在几种类型固定化蛋白质上的应用。
