GSM8403727: KI376_K4me3_starvation_GM07492; Homo sapiens; ChIP-Seq
ChIP-seq in GM07492 fibroblasts were performed during proliferating, at 48 hours into starvation, and at 24 hours into the recovery phase. Cells were crosslinked in 1% formaldehyde for 15 min at room temperature. Cell extract from one million cells was incubated with antibodies in a 200-μL reaction for 12 hours or longer at 4°C. Antibodies used in ChIP are: mouse monoclonal anti-H3K27ac antibody (Wako MABI0309, Lot 14007, RRID:AB_11126964; 2 μL per IP) and mouse monoclonal anti-H3K4me3 antibody (Wako MABI0304, Lot 14004, RRID:AB_11123891; 2 μL per IP). Immunocomplex was captured by Protein G-conjugated magnetic beads and washed. Immunoprecipitated DNA was reverse-crosslinked and used to construct high-throughput sequencing libraries. Immunoprecipitated DNA was reverse-crosslinked and used to construct high-throughput sequencing libraries using NEBNext Ultra DNA Library Prep Kit (New England Biolabs, E7370).