ChIP-Atlas: SRX2378750

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_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: U2OS cells were cross-linked in medium containing 1% formaldehyde for 10 min at room temperature, neutralized with 0.125 M glycine, scraped off and rinsed twice with PBS. Pellets were resuspended in 10 ml of buffer L1 (50 mM Hepes KOH, pH 7.5, 140 mM NaCl, 1 mM EDTA pH 8.0, 10% glycerol, 5% NP-40, 0.25% Triton X-100) and incubated for 10 min at 4°C. Cells were spun down, resuspended in 10 ml of buffer L2 (200 mM NaCl, 1 mM EDTA pH 8.0, 0.5 mM EGTA pH 8.0, 10 mM Tris pH 8.0) and incubated for 10 min at room temperature followed by centrifugation and resuspension in 900 μl buffer L3 (1 mM EDTA pH 8.0, 0.5 mM EGTA pH 8.0, 10 mM Tris pH 8.0, 100 mM NaCl, 0.1% Na-deoxycholate, 0.17 mM N-lauroyl sarcosine, supplemented with protease inhibitors). Library was prepared with Ovation Ultralow Library Systems v2 (NuGEN) according to manufacturer's instructions.