1.6.4 细胞蛋白提取与定量

1.6.4 细胞蛋白提取与定量

去除培养基,用冰PBS洗2次,加入细胞裂解液(25mmol/L Tris·HCl pH 7.6,150mmol/L NaCl,1% NP-40,1% sodium deoxycholate,0.1% SDS,2mmol/L Na3VO4,50mmol/L NaF,4mmol/L Na Pyrophosphate,25μl protein inhibitor),冰上放2min。超声粉碎(1s×3),离心,13000rpm×10min,吸取上清液,弃沉淀。以下步骤与组织蛋白提取后处理相同,见1.3节免疫印迹技术。(https://www.daowen.com)