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文献和实验stirrer.Then 7.5ml of Folin Ciocaltew Reagent was added to each well. The well plate was then left at room temperature for a further 30 minutes. Absorbencies were read at 750nm (A750) in a plate-reading spectrophotometer.A protein standard curve
], and reducing agent [mercaptoenthanol]) are separated by electrophoresis based on their molecular weights, followed by transferring proteins to PVDF membrane and identifying a protein of interest using a specific antibody. It is highly recommended
做了五个月的Western Blot ,有一点点小心得,给新手点经验. 蛋白提取: 1. 总蛋白提取(胞膜,胞浆,胞核): 组织匀浆后,15000g, 4度, 20分钟后,取上清. Buffer:NP40 750ul, 脱氧胆酸钠0.5克, SDS 0.1克, 加1*PBS 至100ml. 再加入PMSF(7.4mg/ml) 0.1ml. (现用现加) 7.4mg/ml PMSP异丙醇溶液: 取A mg PMSF 加 A/7.4 ml 异丙醇.工厂-20度储存 2.组织膜
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