GSM4238562: ChIP-2 SMC3 DLD-1; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
SMC3
Cell type
Cell type Class
Digestive tract
Cell type
DLD-1
Primary Tissue
Colon
Tissue Diagnosis
Adenocarcinoma
Attributes by original data submitter
Sample
source_name
colon epithelium
cell line
epithelial tumor cells DLD-1
cell type
colorectal adenocarcinoma
chip antibody
rabbit polyclonal anti-SMC3, Abcam ab9263
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were fixed at 1% formaldehyde for 10 min at room temperature and quenched with 0.125 mM glycine for 5 min. For some experiments a dual crosslinking was used, by incubating the cells in 1.5 mM ethylene glycol bis(succinimidyl succinate) (EGS) (Sigma) in PBS for 45 min at RT under gentle shaking. Then, 1% formaldehyde was added and the incubation left to proceed for 20 min, after which the fixation was quenched with 0.125 mM glycine for 5 min at RT. Cells were centrifuged, resuspended in PBS/1 x proteinase inhibitor cocktail /10% glycerol, and kept at -80 degrees C. A modified ChIP version named ChEP (chromatin enriched for proteomics) was used to precipitate proteins of interest with specific antibodies. DNA fragments were end-repaired and A-tailed according to Illumina protocol, and then ligated with adapters. DNA fragments that have adapter molecules on both ends were amplified by PCR. DNA concentration of the resulting sequencing libraries was measured with the Qubit 2.0 fluorometer dsDNA HS Assay (Thermo Fisher Scientific), and the size distribution was analyzed using Agilent BioAnalyzer 2100 (Agilent). Paired-end sequencing was performed using an Illumina system with Illumina-provided protocols for paired-end sequencing