Anti h3k27ac antibody

PANC-1 cells (1 × 10⁷) were cross-linked with 1% formaldehyde for 10 min at room temperature. Fixation was stopped by adding 0.125 M glycine. Nuclear pellets were prepared for chromatin immunoprecipitation (ChIP) as described previously (29 (link)). Prewashed magnetic Dynabeads (Life Technologies, MA) were incubated with anti-H3K27ac antibody (Millipore, MA) or anti-SREBP2 antibody (Cayman, MI) in ChIP dilution buffer (20 mM Tris-HCl [pH 8.0], 150 mM NaCl, 1 mM EDTA, 1% Triton X-100, protease inhibitor cocktail [Roche, Basel, Switzerland]) for 6 h by wheel rotating at 4°C. Subsequently, sonicated cross-linked nuclear lysates were added and incubated overnight at 4°C by wheel rotating. The beads were washed several times and eluted with elution buffer (1% SDS and 100 mM NaHCO₃), and the eluent was incubated with pronase (1 mg ml⁻¹) at 42°C for 2 h and then incubated at 65°C overnight. DNA was purified using the QIAquick PCR purification kit (Qiagen, Hilden, Germany). ChIP-seq was done with an Illumina/Solexa sequencer as previously described (29 (link)).