GSM1515524: R1AD1 DHT AR rep2; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
AR
Cell type
Cell type Class
Prostate
Cell type
R1-AD1
NA
NA
Attributes by original data submitter
Sample
source_name
prostate cancer cell line
cell type
R1-AD1
agent
dihydrotestosterone, 1nM
chip antibody
AR -N20 (Santa Cruz, SC-816 lot B1012)
biological replicate number
rep2
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Chromatin immunoprecipitation (ChIP) was performed for three independent biological replicate experiments exactly as described (PMID: 21602788) with the exception fo the following modifications: R1-AD1 and R1-D567 cells (PMID 24101480) were seeded at 5x10^6 cells/plate on 10 cm plates in RMPI 1640 + 10% CSS, allowed to settle for 72h, then re-fed for 4h with RMPI 1640 + 5% CSS containing vehicle (ETH) or 1nM DHT prior to fixation. Nuclear pellets were sonicated on ice for 8 cycles at 40% amplitude using a 450 Sonifer (Branson). Each cycle consisted of 10 sec pulse/10 sec rests for 1 min, with 2 min rests between cycles. Lysates were immunoprecipitated with Protein A/G Plus agarose beads (Santa Cruz Biotechnology) pre-blocked with tRNA (Sigma). DNA was purified using a PCR purification kit (Qiagen). For ChIP-seq, 2-10ng of DNA (ChIP-enriched or input) was used for library creation with a TruSeq ChIP Sample Preparation Kit (Illumina cat #IP-202-1012) and sequenced at the Univeristy of Minnesota Genomics Center using an Illumina HiSeq2000 at 1X50bp. Libraries were prepared according to Illumina's instructions accompanying the TruSeq Sample Preparation Kit (Illumina cat#IP-202-1012).