GSM7680810: Hepatocytes, A1ABDKO, H3K4me1, ChIP, rep2; Mus musculus; ChIP-Seq
Purified hepatocytes were immediately fixed in 1% formaldehyde for 10 minutes. Cells were washed and resuspended in SDS buffer (50 mM Tris pH 8.1, 0.5% SDS, 100 mM NaCl, 5 mM EDTA, and protease inhibitors). The pellet was resuspended in IP buffer (100 mM Tris pH 8.6, 0.3% SDS, 1.7% Triton X-100, and 5 mM EDTA) and sonicated to an average length of 500–1000 bp. Sonicated chromatin was incubated with 10 μg primary antibody (see Suppl.Table 2) overnight, and the immunocomplexes recovered using protein A–conjugated Sepharose beads (Life Tech., 101090). After extensive washes chromatin was de-crosslinked overnight in 0.1 M NaHCO3 and 1% SDS. DNA was purified using PCR purification kit (Qiagen, 28004) Libraries were prepared as described in Blecher-Gonen, R., Barnett-Itzhaki, Z., Jaitin, D. et al. Nat Protoc 8, 539–554 (2013). volume 8 issue 3 pag 539-554.