GSM5103053: JQ1 BRD4 IP rep2; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
BRD4
Cell type
Cell type Class
Blood
Cell type
KBM-7
Primary Tissue
Blood
Tissue Diagnosis
Leukemia Chronic Myelogenous
Attributes by original data submitter
Sample
source_name
BRD4 reporter KBM7 cells (REDS15)
treatment
(+)-JQ1 1 micromolar, 48h
chip-ab
BRD4 (ab128874, Abcam)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
50 million cells were crosslinked with 1% formaldehyde for 10 min at room temperature followed by quenching with 125 mM glycine (pH 2.5) for 5 min on ice. After two times washing with cold PBS, cell pellet was lysed and then the chromatin was sheared using a Covaris S2X (duty cycle: 5%; intensity: 4; power: 10 W; cycles per burst: 200; time: 40 min). The fragmented chromatin was incubated with antibodies overnight at 4 °C, followed by a 3 h incubation with Dynabeads Protein G (Thermal Fisher Scientific) at 4 °C. After washing the beads twice with low-salt RIPA buffer, twice with high-salt RIPA buffer and twice with RIPA-LiCl buffer, the antibody-bound chromatin was eluted, and treated with RNase for 30 min at 37 °C, with proteinase K for 2.5 h at 55°C, and then decrosslinked overnight at 65 °C. DNA was then extracted using phenol-chloroform, precipitated and then dissolved in Tris-EDTA buffer. NEBNext Ultra II DNA Library Prep Kit for Illumina (New England Biolabs, E7645S) with NEBNext Multiplex Oligos for Illumina (New England Biolabs, E7335S and E7500S)