[討論] nuclear extraction
cell membrane 與nuclear membrane成分差不多
都是層脂與一些protein
但是在抽nuclear extraction時
所使用的buffer A B C D
難道就不會破壞核膜嗎?
願聞其詳
謝謝^^
Buffer A: 10 mM HEPES (pH 7.9 at 4oC), 1.5 mM MgCl2 , 10 mM KCL and 0.5 mM
DTT
Buffer B: 0.3 HEPES (pH 7.9), 1.4 M KCL and 0.03 M MgCL2
Buffer C: 20 mM HEPES (pH 7.9), 25% (v/v) glycerol, 0.42 M NaCl, 1.5 mM
MgCl2, 0.2 mM EDTA, 0.5 mM phenylmethylsulfonyl fluoride (PMSF) and 0.5 mM
DTT
Buffer D: 20 mM HEPES (pH 7.9), 20% (v/v) glycerol, 0.1 M KCL, 0.2 mM EDTA,
0.5 mM PMSF, and 0.5 mM DTT. DTT and PMSF were added fresh to the buffers
just before use.
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