Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Histone
Antigen
H2A.ZK4K7K11ac

Cell type

Cell type Class
Prostate
Cell type
VCaP
Primary Tissue
Prostate
Tissue Diagnosis
Carcinoma

Attributes by original data submitter

Sample

source_name
Human cell line
cell type
VCaP (ATCC # CRL-2876). Androgen-sensitive human prostate adenocarcinoma cells
passage number
48
chip antibody
Abcam Cat #ab18262

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ChIP assays were carried out according to the manufacturer’s protocol (Upstate Biotechnology). Briefly, ~ 2 x 10^6 cells were fixed by adding formaldehyde at a final concentration of 1% and incubating for 10 minutes at 37°C. The cells were washed twice with ice cold PBS containing protease inhibitors (1mM phenylmethylsulfonyl fluoride (PMSF), 1 µg/ml aprotinin and 1µg/ml pepstatin A), harvested and treated with SDS lysis buffer for 10 min on ice. The resulting lysates were sonicated to shear the DNA to fragment lengths of 200 to 500 basepairs. The complexes were immunoprecipitated with antibodies specific for Histone H2AZ (Active motif # 39113), acetylated Histone H2AZ (Abcam #ab18262), H3K4Me1 (Active Motif #39297), H3K27ac (Active Motif #39133), H3K9K14ac (Millipore #06-599.) and H3K36Me3 (Abcam ab9050-100). Ten µl of antibody was used for each immunoprecipitation. No antibody controls were also included for each ChIP assay and no precipitation was observed by quantitative Real-Time PCR (qPCR) analysis. Input samples were processed in parallel. The antibody/protein complexes were collected by either salmon sperm DNA/protein A agarose slurry or Protein A/G PLUS agarose beads (Santa Cruz sc-2003) and washed several times. The immune complexes were eluted with 1% SDS and 0.1 M NaHCO3 and samples were treated with proteinase K for 1 hour and DNA was purified by phenol/chloroform extraction, ethanol precipitation and resuspended in 30 ul H2O. TruSeq ChIP sample prep kit

Sequencing Platform

instrument_model
NextSeq 500

hg38

Number of total reads
27245755
Reads aligned (%)
98.3
Duplicates removed (%)
11.0
Number of peaks
12613 (qval < 1E-05)

hg19

Number of total reads
27245755
Reads aligned (%)
97.4
Duplicates removed (%)
12.5
Number of peaks
12495 (qval < 1E-05)

Base call quality data from DBCLS SRA