ChIP-Atlas: SRX1023937

Antigen

Cell type Class: Blood
Cell type: Monocytes-CD14+
Tissue: monocytes
Lineage: mesoderm
Description: Monocytes-CD14+ are CD14-positive cells from human leukapheresis production, from donor RO 01746 (draw 1 ID is RO 01746, draw 2 ID is RO 01826), newly promoted to tier 2: not in 2011 analysis

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: In order to perform ChIP experiments the chromatin of day 11 cells was cross-linked for 8 min with 1% formaldehyde in PBS. Then the formaldehyde was quenched for 5 min with 125 mM of glycine. Nuclei were isolated and the chromatin was sheared with 30 cycles at high intensity (30 sec off and 30 sec on) using a sonicator (BioruptorTM Next Gene, Diagenode). For each immunoprecipitation 5 μg of sheared chromatin was used. Pre-cleared chromatin was incubated overnight with 5 μg of an antibody against the active enhancer mark H3K27ac (Abcam; ab4729). Following day the antibodies were captured with 25 μl of protein A magnetic beads (Millipore)s for 2 hours on a rotating wheel at 4°C. The DNA was purified with a QIAquick PCR clean-up kit. ChIP sequencing was performed at the GeneCore facility of EMBL Heidelberg (http://www.genecore.embl.de/) according to the Illumina protocol.