全蛋白提取试剂盒（中）
储存条件	-20℃
单位	盒
全蛋白提取试剂盒（中）用于哺乳动物组织和细胞中提取总蛋白，试剂盒中的裂解液含有蛋白酶抑制剂和磷酸酶抑制剂，作用较为温和，可快速获得总蛋白，可用于免疫印迹实验等基础研究实验，由于含有上述的酶抑制剂，不能用于研究蛋白激酶和磷酸激酶的研究。本品仅用于科研。
产品组成:

操作方法:
1、组织总蛋白的提取
在1ml的冷裂解液中各加入10ul的磷酸酶抑制剂、蛋白酶抑制剂和PMSF；混匀，放置在冰上备用；称取0.1g的新鲜组织，放置于玻璃匀浆器的入口缓冲处，用眼科剪将组织块尽可能的剪碎，然后加入0.5-1ml的新配置的裂解液，研磨直至没有明显的组织块，这个过程注意冰上操作；将组织匀浆液移入1.5ml的EP管中，4 oC 12000g离心30min；吸取上清至新的管中；进行蛋白定量或者变性进行蛋白实验。（提取好的蛋白建议保存于-80 oC，即用即取，避免反复冻融，避免长时间储存。)
2、细胞总蛋白的提取
裂解液的用量：107个细胞需要裂解液1ml；
贴壁细胞：弃去培养基，用冷的PBS洗两次，弃去PBS，然后加入计算好的细胞裂解液；在冰上用细胞刮刀进行刮离细胞，将刮离的细胞裂解液移入EP管中，颠倒裂解20-30min。
悬浮细胞：4 oC 400g离心收集细胞，用冷的PBS洗两次细胞，同样按细胞数加入裂解液，涡旋10s，放置冰上裂解10min，重复3-4次；裂解完毕之后，将细胞裂解液4 oC 12000g离心30min；将上清移入新的EP管中；进行蛋白定量或者变性进行蛋白实验。
（提取好的蛋白建议保存于-80 oC，即用即取，避免反复冻融，避免长时间储存。)
注意事项：
实验过程，所有试剂都要需要预冷或者即融，保证操作过程中的低温环境；
PSMF（有毒）现用现加，因为PMSF在水溶液中会快速降解；
 
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