HeLa细胞裂解+细胞核提取+分离纯化线粒体
一、细胞的裂解HeLa cells were pelleted using a Sorvall centrifuge at 800 g for 10 min.2. Cells were resuspended in lysis buffer [50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.5% NP-40, 5 mM EDTA, 50 mM NaF, mM PMSF and a protease inhibitor(PI) cocktail: 25 μg/ml pepstain A, 50 μg/ml leupeptin and 0.2% aprotinin] and allowed to swell on ice for 30 min.3. Cells were then frozen in a dry ice bath and thawed on ice.4. Cells were spun at 10 000 rpm for 10 min and the supernatant was frozen at -80℃.5. Bradford reagent was used to quantitate the protein concentration per sample.二、提取细胞核Cells were sedimented at 800 g for 10 min and then washed with 50 vol of phosphate-buffered saline(PBS)