ChIP-Atlas: SRX5086087

Antigen

Cell type Class: Blood
Cell type: Th2 Cells
MeSH Description: A subset of helper-inducer T-lymphocytes which synthesize and secrete the INTERLEUKINS IL-4; IL-5; IL-6; and IL-10. These cytokines influence B-cell development and antibody production as well as augmenting humoral responses.

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: Cells (10 x 10e6) were cross-linked for 10 min with 1% formaldehyde, quenched with Glycine, and washed. Cell pellets were resuspended in lysis buffer, and homogenized. Nuclei were resuspended in RIPA buffer and sonicated to a size range of 200-500 bp using Bioruptor. Cells were pelleted, RIPA buffer was added to supernatant to 1 ml, and appropriate antibody coupled to Dynabeads M-280 Sheep anti-rabbit IgG was added. The immunocomplexes were washed, supernatants were collected, incubated at 65oC overnight, and immunoprecipitated DNA were purified using MinElute columns (Qiagen). Antibodies: Rabbit anti-BATF (Schraml et al., 2010), rabbit anti-CTCF (Active Motif, 61311), rabbit anti-Med12 (Bethyl, A300-774A), rabbit anti-Ets1 (Santa Cruz, sc 350X), rabbit anti-IRF4 (Santa Cruz, sc-28696 X), and normal rabbit IgG (Millipore, 12-370). Immunoprecipitated DNA was used to prepare library using NEBNext Ultra II DNA library prep kit and NEBNext Multiplex Oligos for Illumina (NEB). ChIP-seq libraries were sequenced with single-end 50 bp using Illumina HiSeq 2500.