THE BINDING INTERACTION OF COOMASSIE BLUE WITH PROTEINS
THE BINDING INTERACTION OF COOMASSIE BLUE WITH PROTEINS
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DOI：
10.1006/abio.1993.1439
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发表时间：
1993-09-01
影响因子：
2.9
通讯作者：
SPLITTGERBER, AG
中科院分区：
文献类型：
--
作者：
CONGDON, RW;MUTH, GW;SPLITTGERBER, AG
考马斯亮蓝蛋白测定法由于其灵敏度和方便性而被普遍使用，但在分子水平上还没有得到很好的理解。本研究试图通过在正常蛋白质测定的溶剂条件下对选定蛋白质进行分光光度结合研究，更好地了解测定系统。研究通常在高蛋白/染料浓度比下进行，其中仅高亲和力染料结合位点将被占据。修改后的Scatchard和希尔分析表明，这些高亲和力网站的数量很少相比，染料结合位点的总数在一个给定的蛋白质。高亲和力结合常数的大小是典型的非共价结合相互作用。
The Coomassie brilliant blue protein assay is commonly used because of its sensitivity and convenience, but it is not well understood on a molecular level. This study attempts to gain better understanding of the assay system through spectrophotometric binding studies carried out on selected proteins under solvent conditions characteristic of the normal protein assay. The studies were generally conducted at high protein/dye concentration ratios, where only the high-affinity dye binding sites would be occupied. Modified Scatchard and Hill analyses show that these high-affinity sites are few in number compared to the total number of dye-binding sites on a given protein. The magnitudes of the high-affinity binding constants are typical of noncovalent binding interactions.