使用说明：
1.  准备溶液：室温融解试剂盒中的三种试剂，溶解后立即放置在冰上，混匀。取适当量的细胞浆蛋白
    抽提试剂A备用，在使用前数分钟内加入PMSF，使PMSF的最终浓度为1mM。取适当量的细胞核蛋
    白抽提试剂备用，在使用前数分钟内加入PMSF，使PMSF的最终浓度为1mM。
2.  对于贴壁细胞：用PBS洗一遍，用细胞刮子刮下细胞，或用EDTA溶液处理细胞使细胞不再贴壁很
    紧，并用移液器吹打下细胞。离心收集细胞，尽最大努力吸尽上清，留下细胞沉淀备用。尽量避免
    用胰酶消化细胞，以免胰酶降解需抽提的目的蛋白。
3.  对于悬浮细胞：用PBS洗一遍，离心收集细胞，尽最大努力吸尽上清，留下细胞沉淀备用。
4.  每20微升细胞沉淀加入200微升添加了PMSF的细胞浆蛋白抽提试剂A。(对于二百万Hela细胞，其细
    胞沉淀的体积大约为20微升或40毫克。)
5.  最高速剧烈Vortex 5秒，把细胞沉淀完全悬浮并分散开。(如果细胞沉淀没有完全悬浮并分散开，可
    以适当延长vortex时间。)
6.  冰浴10-15分钟。
7.  加入细胞浆蛋白抽提试剂B 10微升。最高速剧烈Vortex 5秒，冰浴1分钟。
8.  最高速剧烈Vortex 5秒，4℃ 12,000-16,000g离心5分钟。
9.  立即吸取上清至一预冷的塑料管中，即为抽提得到的细胞浆蛋白。可以立即使用，也可以冻存。(千
    万不要触及沉淀，可以在沉淀上方保留极小体积的上清，以免触及沉淀。每200万细胞用200微升本
    产品中的细胞浆蛋白抽提试剂A裂解后可获得的上清，其细胞浆蛋白浓度约为2-5mg/ml，不同细胞有
    所不同。)
10. 对于沉淀，完全吸尽残余的上清，加入50微升添加了PMSF的细胞核蛋白抽提试剂。(不吸尽上清会
    带来细胞浆蛋白的污染。)
11. 最高速剧烈Vortex 15-30秒，把细胞沉淀完全悬浮并分散开。然后放回冰浴中，每隔1-2分钟再高速
    剧烈Vortex 15-30秒，共30分钟。
12. 4℃ 12,000-16,000g离心10分钟。
13. 立即吸取上清至一预冷的塑料管中，即为抽提得到的细胞核蛋白。可以立即使用，也可以-70℃冻
    存。每200万细胞用50微升本产品中的细胞核蛋白抽提试剂裂解后获得的上清，其细胞核蛋白浓度约
    为1.2-3.0mg/ml，不同细胞有所不同。
14. 对于新鲜组织：
    a. 把组织尽可能切成非常细小的碎片。按照20:1的比例混合适当量的细胞浆蛋白抽提试剂A和B(例
       如200微升细胞浆蛋白抽提试剂A中加入10微升抽提试剂B)。并加入PMSF至最终浓度为1mM配
       制成组织匀浆液。按照每60mg组织加入200微升组织匀浆液的比例混合组织和组织匀浆液，并在
       玻璃匀浆器内充分匀浆。匀浆需在冰浴或4℃进行。
    b. 匀浆后把匀浆液转移到塑料离心管内，冰浴放置15分钟。
    c. 4℃ 1,500g离心5分钟。把上清转移至一预冷的塑料管中，为抽提得到的部分细胞浆蛋白。(吸上
       清时千万不要触及沉淀。)
    d. 对于沉淀，到了这一步已经充分匀浆，但很多细胞还没有破碎。接下去按照使用说明的步骤4开
       始操作，即把沉淀当做已经离心收集好的细胞沉淀操作，按照步骤4至步骤13抽提得到细胞浆蛋
       白和细胞核蛋白。抽提得到的细胞浆蛋白可以和步骤14c中抽提得到的细胞浆蛋白合并。每60mg
       新鲜的肝脏组织用本产品裂解后获得的上清，其细胞浆蛋白浓度约为3-10mg/ml，细胞核蛋白浓
       度约为3-10mg/ml，不同状态的不同组织有所不同。

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