characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-03-14T17:01:19Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2016-01-28T12:20:33Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA3859819
characteristics.characteristic: External Id
characteristics.text: Research Associate
characteristics.characteristic: INSDC center name
characteristics.text: 2017-03-14T17:01:19Z
characteristics.characteristic: INSDC first public
characteristics.text: 2016-01-28T12:20:33Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1046953
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-4358:TCF3-IgG
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: embryonic stem cell
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0002322
characteristics.characteristic: cell type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: 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.
characteristics.characteristic: description
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: TCF3-IgG
characteristics.characteristic: title