SAMPLE COLLECTION AND STORAGE

Cell Lysates - Cells must be lysed before assaying according to the following directions.

1.  Adherent cells should be detached with trypsin and then collected by centrifugation  (suspension cells can be collected by centrifugation directly).

2.  Wash cells three times in cold PBS.

3.  Resuspend cells in PBS (1×) and the cells was subject to ultrasonication for 4 times (or Freeze cells at ≤-20^oC. Thaw cells with gentle mixing. Repeat the freeze/thaw cycle for 3 times.)

细胞裂解液：

   1）贴壁细胞需要先用胰酶消化，离心收集细胞（悬浮细胞可直接离心收集）；

   2）将收集到的细胞用冷PBS洗3次；

   3）物理方法裂解细胞（可先超声破碎细胞，再反复冻融）：

      i 超声破碎：取适量PBS重悬细胞，用一定功率的超声波处理细胞悬液，使细胞急剧震荡破裂。

      ii 反复冻融：将待破碎的细胞在-20^oC以下冰冻，室温融解，反复3次，使细胞溶胀破碎。

   4）将标本于2-8^oC 1500×g离心10分钟，收集上清备用。