Illumina HiSeq 2500 sequencing; Identification of target genes of the transcription factors TCF1 and TCF3 in mouse embryonic stem cells (ChIP-Seq)
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
No description
Antigen
NA
Cell type
Cell type Class
Pluripotent stem cell
Cell type
ES cells
NA
NA
Attributes by original data submitter
Sample
ENA first public
2017-03-14
ENA last update
2016-01-28
ENA-CHECKLIST
ERC000011
External Id
SAMEA3859817
INSDC center alias
Research Associate
INSDC center name
Research Associate
INSDC first public
2017-03-14T17:01:19Z
INSDC last update
2016-01-28T12:20:33Z
INSDC status
public
Submitter Id
E-MTAB-4358:TCF1-IgG
broker name
ArrayExpress
cell type
embryonic stem cell
common name
house mouse
sample name
E-MTAB-4358:TCF1-IgG
Sequenced DNA Library
library_name
TCF1-IgG
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
R1 and E14Tg2 mouse ESCs were cultured on 0,1% gelatin in ESC medium: knock-out DMEM supplemented with 15% FBS (Sigma) and 1,000 U/ml LIF ESGRO (Millipore). mESCs were treated at indicated concentrations to activate the Wnt pathway: purified Wnt3a (Peprotech); BIO (Calbiochem); CHIR99021 (Calbiochem). BIO and CHIR99021 were resuspendend in DMSO (Sigma). Briefly, ESCs were trypsinised and crosslinked in 1% formaldehyde for 10 min at room temperature. Crosslinking was quenched with 0.125 M glycine for 5 min. The pelleted cells were lysed in 1 ml ChIP buffer and sonicated for 10 min in a Bioruptor sonicator (Diagenode). The soluble material was quantified using Bradford assays. To immunoprecipitate the transcription factors, 500 g protein was used. Antibodies were incubated overnight with the chromatin. The immunocomplexes were recovered with 30 l protein A or G agarose bead slurries. The immunoprecipitated material was washed three times with low-salt buffer and one time with high-salt buffer.