Cold Spring Harb Protoc. 2020 Feb 3;2020(2):098582. doi: 10.1101/pdb.prot098582.
Differential detergent fractionation of cells is a rapid method for extraction of cytoplasmic and nuclear proteins in preparation of an immunoprecipitation. This method can be applied for use of adherent or suspension cells and can significantly reduce nonspecific background in an immunoprecipitation by separation of cellular compartments into individual fractions. The lysis of cells by differential detergents permits the rapid extraction of proteins from the cytoplasm (digitonin), the cytoplasmic membranes, and organelles (Triton X-100), and nucleoplasm (Tween/DOC), facilitated through the use of distinct extraction buffers. Cytoplasmic and nuclear matrix proteins as well as DNA are left behind during the detergent-based extraction.
细胞差异去污剂分步分离是一种快速提取细胞质和核蛋白以准备免疫沉淀的方法。该方法可用于贴壁或悬浮细胞，通过将细胞区室分离成单独的级分，可以显着降低免疫沉淀中的非特异性背景。通过使用不同的提取缓冲液，差异去污剂裂解细胞可从细胞质（二氢月桂酰基牛磺酸钠）、细胞质膜和细胞器（曲拉通 X-100）以及核质（吐温/十二烷基硫酸钠）中快速提取蛋白质。细胞质和核基质蛋白以及 DNA 在基于去污剂的提取过程中被留下。
