Harlow Ed, Lane David
CSH Protoc. 2006 Sep 1;2006(4):pdb.prot4531. doi: 10.1101/pdb.prot4531.
INTRODUCTIONFor cells grown in tissue culture, the most useful method of lysis is treating with detergents, as described in this protocol. Non-ionic detergents, such as NP-40, solubilize the plasma and intracellular membranes, break many weak intermolecular bonds, and solubilize most of the commonly studied protein antigens. RIPA lysis buffer, which contains non-ionic detergents mixed with ionic detergents, may be used as a more rigorous extraction buffer to release all but the insoluble proteins of the cell and to break most weak noncovalent interactions. The resulting lysate is ready for preclearing.
引言
对于在组织培养中生长的细胞，如本方案所述，最有用的裂解方法是用去污剂处理。非离子去污剂，如NP - 40，可溶解质膜和细胞内膜，破坏许多弱分子间键，并溶解大多数常用研究的蛋白质抗原。含有非离子去污剂与离子去污剂混合的RIPA裂解缓冲液，可用作更严格的提取缓冲液，以释放细胞中除不溶性蛋白质外的所有蛋白质，并打破大多数弱非共价相互作用。所得裂解物可用于预清除。
