ChIP-Atlas: SRX4457302

Antigen

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: Nascent RNA capture was performed using Click-iT® Nascent RNA Capture Kit according to the manufacturer's instructions with minor modifications. RIP-seq libraries:H2A.Z-; acH2A.Z-; TIP60-; and IgG-interacting RNA was immunoprecipitated by the respective antibodies. RNA was reverse transcribed into ds-cDNA before sequencing library preparation using Illumina ChIP-Seq sample preparation kit (cat# IP-102-1001) following manufacturer's instruction. ChIP-seq libraries: Cells were crosslinked with 1% formaldehyde for 10 min at 37°C. Pellets of 1M cells were used for immunoprecipitation as previously described (7) and lysed for 10 minutes on ice and chromatin fragmented using a Branson 250 digital sonifier. Each ChIP was performed with 4ug of antibody, incubated overnight at 4°C. A 50/50 slurry of protein A and protein G Dynabeads was used to capture enriched chromatin, which was then washed before reverse-crosslinking and proteinase K digestion at 65C. AMPure XP beads were used to clean up and isolate ChIP DNA for subsequent library construction. The following antibodies were used for ChIP: H2A.Z (Abcam ab4174, lot GR3176820-1), acH2A.Z (Abcam ab18262, lot GR306397-1), and IgG (ab171870).