title: TCF1-IgG
organism: Mus musculus
description: 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.
attributes.tag: ENA first public
attributes.value: 2017-03-14
attributes.tag: ENA last update
attributes.value: 2016-01-28
attributes.tag: External Id
attributes.value: SAMEA3859817
attributes.tag: INSDC center alias
attributes.value: Research Associate
attributes.tag: INSDC center name
attributes.value: Research Associate
attributes.tag: INSDC first public
attributes.value: 2017-03-14T17:01:19Z
attributes.tag: INSDC last update
attributes.value: 2016-01-28T12:20:33Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-4358:TCF1-IgG
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: embryonic stem cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: sample name
attributes.value: E-MTAB-4358:TCF1-IgG