Stoscheck C M
Department of Medicine, Veterans Administration and Vanderbilt Medical Centers, Nashville, Tennessee 37212.
Anal Biochem. 1990 Jan;184(1):111-6. doi: 10.1016/0003-2697(90)90021-z.
Coomassie blue G dye-based protein assays are exceptionally convenient because of their simplicity, sensitivity, speed, and resistance to interfering chemicals, notably reducing agents and most buffers. A major problem with the assay is the variation in response to different proteins. The addition of NaOH to the protein assay reagent reduced the variation in the response of this assay to different proteins. In addition, the sensitivity of the assay is increased. The NaOH can be added either in a separate step to solubilize cells or membranes or directly to the reagent. Linear standard curves were obtained when the log of the absorbance was plotted against the log of the protein quantity.
基于考马斯亮蓝G染料的蛋白质测定法非常方便,因为其具有简单、灵敏、快速以及抗干扰化学物质(尤其是还原剂和大多数缓冲液)的特点。该测定法的一个主要问题是对不同蛋白质的反应存在差异。在蛋白质测定试剂中加入氢氧化钠可减少该测定法对不同蛋白质反应的差异。此外,测定法的灵敏度也会提高。氢氧化钠既可以在单独的步骤中添加以溶解细胞或膜,也可以直接添加到试剂中。当将吸光度的对数与蛋白质量的对数作图时,可得到线性标准曲线。