Tsuruoka Metabolomics Laboratory, National Cancer Center, Tsuruoka, Yamagata 997-0052, Japan.
Division of Hematological Malignancy, National Cancer Center Research Institute, Chuo-ku, Tokyo 104-0045, Japan.
STAR Protoc. 2021 Mar 23;2(2):100404. doi: 10.1016/j.xpro.2021.100404. eCollection 2021 Jun 18.
Conventional chromatin immunoprecipitation (ChIP) includes many steps that need to be optimized. Here, we have described a protocol of fractionation-assisted native ChIP (fanChIP) which combines subfractionation and native ChIP to purify protein/chromatin complexes applicable for analyses of both protein-protein and protein-DNA interactions within a short period of time. fanChIP is advantageous as subcellular fractionation removes chromatin-unbound materials before immunoprecipitation, and the chromatin fragmentation by micrococcal nuclease (MNase) in a mild condition enables one-step purification of intact protein/chromatin complexes. For complete details on the use and execution of this protocol, please refer to Miyamoto et al. (2020).
常规染色质免疫沉淀（ChIP）包括许多需要优化的步骤。在这里，我们描述了一种分馏辅助天然染色质免疫沉淀（fanChIP）的方案，该方案将亚细胞分馏与天然 ChIP 相结合，以纯化适用于短时间内分析蛋白质/DNA 相互作用的蛋白质/染色质复合物。fanChIP 的优点是亚细胞分馏在免疫沉淀前去除了非染色质结合的物质，而微球菌核酸酶（MNase）在温和条件下的染色质片段化使得完整的蛋白质/染色质复合物可以一步纯化。有关此方案使用和执行的完整详细信息，请参阅 Miyamoto 等人（2020 年）。
