试剂准备 cold lysis buffer:(胞质蛋白提取裂解液) RNA structure buffer:
1mM PMSF 10 mM Tris pH 7.0
20units Rnasin 0.1 M KCl
10mM Tris-HCl pH 7.4 10 mM MgCl2
10mM NaCl
3mM MgCl2
0.1mM DTT
0.5% NP40
RIP buffer:(胞核蛋白裂解液) nuclear isolation buffer(核提取液):
150 mM KCl 1.28 M sucrose
25 mM Tris pH 7.4 40 mM Tris-HCl pH 7.5
0.5 mM DTT 20 mM MgCl2
0.5% NP40 4% Triton X-100
1 mM PMSF
protease Inhibitor