GSM7383795: T cells, BATF-TFAP4 knockin, BATF antibody, donor 2; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
BATF
Cell type
Cell type Class
Blood
Cell type
T cells
NA
NA
Attributes by original data submitter
Sample
source_name
Blood
individual
donor 2
tissue
Blood
cell line
Primary cells
cell type
T cells
genotype
BATF-TFAP4 knockin
treatment
None
chip antibody
BATF
geo_loc_name
missing
collection_date
missing
Sequenced DNA Library
library_name
GSM7383795
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Sorted CAR T cells (5e6) were double crosslinked by 50 mM DSG (disuccinimidyl glutarate, ProteoChem) for 30 min followed by 10 min of 1% formaldehyde. Formaldehyde was quenched by the addition of glycine. Nuclei were isolated using ChIP lysis buffer (1% Triton x-100, 0.1% SDS, 150 mM NaCl, 1 mM EDTA, and 20 mM Tris, pH 8.0). Nuclei were sheared with Covaris sonicator using the following setup: Fill level – 10, Duty Cycle – 5, PIP – 140, Cycles/Burst – 200, Time – 4 min. Sheared chromatin was immunoprecipitated overnight with the following antibodies: BATF (brookwoodbiomedical – pab4003), TFAP4 (kind gift from Takeshi Egawa's group). Antibody chromatin complexes were pulled down with Protein A magnetic beads and washed once in IP wash buffer I. (1% Triton, 0.1% SDS, 150 mM NaCl, 1 mM EDTA, 20 mM Tris, pH 8.0, and 0.1% NaDOC), twice in IP wash buffer II. (1% Triton, 0.1% SDS, 500 mM NaCl, 1 mM EDTA, 20 mM Tris, pH 8.0, and 0.1% NaDOC), once in IP wash buffer III. (0.25 M LiCl, 0.5% NP-40, 1mM EDTA, 20 mM Tris, pH 8.0, 0.5% NaDOC) and once in TE buffer (10 mM EDTA and 200 mM Tris, pH 8.0). DNA was eluted from the beads by vigorous shaking for 20 minutes in elution buffer (100 mM NaHCO3, 1% SDS). DNA was decrosslinked overnight at 65C and purified with MinElute PCR purification kit (Qiagen). DNA was quantified by Qubit and 10 ng DNA was used for sequencing library construction with the Ovation Ultralow Library System V2 (Tecan) using 12 PCR cycles according to the manufacturer's recommendations.