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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.)

細(xì)胞裂解液：

   1）貼壁細(xì)胞需要先用胰酶消化，離心收集細(xì)胞（懸浮細(xì)胞可直接離心收集）；

   2）將收集到的細(xì)胞用冷PBS洗3次；

   3）物理方法裂解細(xì)胞（可先超聲破碎細(xì)胞，再反復(fù)凍融）：

      i 超聲破碎：取適量PBS重懸細(xì)胞，用一定功率的超聲波處理細(xì)胞懸液，使細(xì)胞急劇震蕩破裂。

      ii 反復(fù)凍融：將待破碎的細(xì)胞在-20^oC以下冰凍，室溫融解，反復(fù)3次，使細(xì)胞溶脹破碎。

   4）將標(biāo)本于2-8^oC 1500×g離心10分鐘，收集上清備用。