ChIP-Atlas: ERX519498

Antigen

Cell type Class: Bone
Cell type: U2OS
Tissue: bone
Lineage: mesoderm
Description: osteosarcoma from the tibia of a 15 year old, J. Ponten and E. Saksela derived this line (originally 2T) in 1964 from a moderately differentiated sarcoma, viruses were not detected during co-cultivation with WI-38 cells or in CF tests against SV40, RSV or adenoviruses, mycoplasma contamination was detected and eliminated in 1972, (PMID: 6081590)

library_name: GR_gamma
library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: Approximately 10 million cells were treated with 0.01% ethanol vehicle or 1 uM dexamethasone for 1.5 h. Chromatin was sheared with a Bio-ruptor water bath sonicator (Diagenode) to produce fragments of 100-200 bp. Protein-G coupled magnetic beads (Active- Motif) were preincubated for 1 h with GR-antibody (N499) before chromatin was added and incubated for an additional 2-4 h while rotating at 4C. Subsequently beads were washed three times with 10 mM TrisaHCl pH 8.0, 1 mM EDTA, 500 mM NaCl, 5% (vol/vol) glycerol, 0.1% sodium deoxycholate, 0.1% SDS, 1% Triton X-100, 0.5 mg/uL BSA, followed by three additional washes with 20 mM Tris, pH 8.0, 1 mM EDTA, 250 mM LiCl, 0.5% Nonidet P-40, and 0.5% sodium deoxycholate.