GSM5553101: 42D-ENZR-ASCL1-ChIP; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
ASCL1
Cell type
Cell type Class
Prostate
Cell type
LNCAP
Primary Tissue
Prostate
Tissue Diagnosis
Carcinoma
Attributes by original data submitter
Sample
source_name
42D Prostate cancer cell line generated from mouse xenograft of LNCaP
cell type
ENZ-resistant CRPC-NE
treatment
Enzalutamide
shRNA
AR positive, Neuro Endocrine like
antibody
ASCL1
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cell lines were grown in media supplemented with 5% FBS (Gibco #A3160701) and processed for ChIP using the Magna ChIP Kit (Millipore #17-10085) according to manufacturer's instructions with the following antibodies: ASCL1 (5 µg, Abcam#556604, EZH2 (5 μg, Active Motif #39933), H3K27Me3 (5 µg; Millipore #07-449). 10 million cells were fixed with 1% formaldehyde for 10 minutes at room temperature. Cells were collected. Chromatin was sonicated to approximately 500bp. For ChIP-seq, sequencing libraries (100 ng DNA per sample) were constructed using the KAPA library Prep Kit (Roche, Cat KK8510) with Illumina TruSeq indexes. Libraries were assessed for quality using gel electrophoresis and libraries passing quality control (e.g., no primer dimers) were quantified using the KAPA Library Quantification Kit for Illumina (Roche, Cat KK4824). Libraries were sequenced using the Illumina NextSeq 500 (75 bp single-end reads; cell lines).