ChIP-Atlas: ERX1030597

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: ZJ1_H3K18ac
library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: 'The human osteosarcoma U2OS cells were grown in DMEM supplement with 10% foetal bovine serum for 24h, then cells were transfected for 48 hours with ON-TARGETplus Non-targeting Pool (Dharmacon D-001810-10) using Lipofectamine RNAiMax (Life technologies) according to manufacturer''s protocol. Cells were cross-linked with 1% Formaldehyde for 10 minutes.' After reverse cross-link and protein digestion with Proteinase K (Roche), DNA was purified using Qiaquick, PCR purification kit (Qiagen) following the manufacturer’s instructions. ChIP-seq was performed as described in Schmidt et al., 2009, using 10 x 106 U2OS. Immuno-precipitation was performed overnight at 4° C by incubating the shared DNA-protein complex with 10 μg of anti-H3K18Ac (Abcam AB1191 ) antibodies previously coupled to Dynabeads protein G (Invitrogen). Schmidt D, Wilson MD, Spyrou C, Brown GD, Hadfield J, Odom DT. (2009) ChIP-seq: using high-throughput sequencing to discover protein-DNA interactions. Methods 48: 240–248.