Department of Environmental Biosciences, Yokohama City University, Kanagawa, Japan.
Anal Biochem. 2010 Nov 1;406(1):86-8. doi: 10.1016/j.ab.2010.06.035. Epub 2010 Jun 26.
Coomassie Brilliant Blue (CBB) protein stains are inexpensive but detect proteins at only at microgram levels. Because of acetic acid and methanol, they cause skin irritation and reduce work motivation by malodor. Recent mass spectrometric (MS) analyses demonstrated that nanogram-sensitive colloidal CBB staining resulted in in vitro methylations of proteins. We propose a rapid, inexpensive, sensitive, odorless, less harsh, and in vitro methylation-free CBB stain. CGP uses three components: citric acid, CBB G-250, and polyvinylpyrrolidone. CGP detects proteins at 12ng within 45min, and because it is nonalcohol, in principle in vitro methylation would be eliminated. Indeed, MS analysis of CGP-stained bands confirmed a lack of methylation.
考马斯亮蓝(CBB)蛋白染色剂价格便宜,但只能检测微克水平的蛋白质。由于含有乙酸和甲醇,它们会引起皮肤刺激,并通过恶臭降低工作动力。最近的质谱(MS)分析表明,纳克敏感的胶态 CBB 染色导致蛋白质的体外甲基化。我们提出了一种快速、廉价、敏感、无臭、温和且无体外甲基化的 CBB 染色剂。CGP 使用三种成分:柠檬酸、CBB G-250 和聚乙烯吡咯烷酮。CGP 在 45 分钟内可检测到 12ng 的蛋白质,而且由于它不含酒精,理论上可以消除体外甲基化。事实上,MS 分析 CGP 染色条带证实不存在甲基化。