GSM4194716: RPE1 Input-interphase-ChIP-seq-r3; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Input control
Antigen
Input control
Cell type
Cell type Class
Neural
Cell type
HRPEpiC
Tissue
epithelium
Lineage
ectoderm
Description
retinal pigment epithelial cells
Attributes by original data submitter
Sample
source_name
RPE1 retinal pigment epithelial cell line
cell line
RPE1
cell cycle phase
Interphase
replicate
3
antibody
none
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were cross-linked with 1% formaldehyde at room temperature for 10 min and neutralized with 0.125 M glycine for 5 min . After sonication, 25 ug of chromatin was incubated with 5 μg of anti-H3K4me3, anti-H3K4me1, anti-H3K27ac, or anti-CTCF antibodies at 4°C overnight. For spike-in normalization, 50 ng of Spike-in chromatin (53083, Active Motif) with 2 μg Spike-in antibody were added. Immunoprecipitated complexes were collected using Dynabeads M280 sheep-anti-rabbit IgG (Invitrogen). Subsequently, immuno-complexes were washed, decorsslinked and DNA was purified by QIAquick Spin columns (Qiagen). Cells (5 × 106) were cross-linked with 1% formaldehyde for 10 min, and ChIP-seq was performed as previously described (Toyama et al. 2018). Spike-in was carried out according to vendor protocols (Active Motif). Briefly, DNA libraries were generated using the Kapa Hyper Prep Kit for Illumina Platforms (Kapa biosystems). Libraries were sequenced in a NextSeq 500 system (Illumina). ChIP-seq libraries were generated using the Kapa Hyper Prep Kit for Illumina Platforms (Kapa biosystems).