Sample Detail

TitleE-MTAB-1851:Hd-S4
DescriptionProtocols: The preparation of the material was as described in Elsby et al., 2009. HeLa cells were crosslinked and frozen in order to pool enough for ChIP. Elsby,L.M., Donn,R., Alourfi,Z., Green,L.M., Beaulieu,E. and Ray, D.W. (2009) Hypoxia and glucocorticoid signaling converge to regulate macrophage migration inhibitory factor gene expression. Arthritis Rheum., 60, 2220-2231. ChIP assays were performed as described in Elsby et al., 2009, with the following modifications for ChIP-seq. The crosslinked material was sheared by twice probe sonication (SONICS VCX130) with energy more than 50% as 12 secs on/ 40secs off, followed by water sonication using bio-rupter (Next Generation) for 40 mins, at high level, 30 secs on/ 30 secs off, 4 degrees C. The size of sheared DNA fragments was between 100 bp to 500 bp, with most was around 200 bp. Immunoprecipitation was performed by incubating 1x10^7cells with 3 ug of GR antibodies (1ug anti-GR M-20 and 1 ug anti-GR H-300X from Santa Cruz Biotechnology, and 1 ug anti-GR from Sigma-Aldrich), overnight at 4 degrees C, followed by protein G dynabeads (Invitrogen). Elsby,L.M., Donn,R., Alourfi,Z., Green,L.M., Beaulieu,E. and Ray, D.W. (2009) Hypoxia and glucocorticoid signaling converge to regulate macrophage migration inhibitory factor gene expression. Arthritis Rheum., 60, 2220-2231.

Organism Info

Taxon ID9606
Common Name
Scientific NameHomo sapiens
Anonymized Name
Individual Name