GSM4388159: THP1_IgG_rep2; Homo sapiens; OTHER
20 million MOLM-13 and THP-1 cells were UV crosslinked at 400 mJoules/cm2 with 254 nm radiation, and snap frozen. Cells were then lysed and treated with RNase I to fragment RNA as previously described. Anti-ZFP36L2/TIS11D antibody (ab70775) was then pre-coupled to Sheep anti-Rabbit Dynabeads (Thermo Fisher), added to lysate, and incubated overnight at 4 deg C. Prior to immunoprecipitation, 2% of the sample was taken as the paired input sample, while the remainder was magnetically separated and processed using the standard eCLIP protocol. eCLIP was performed by excising the area from ~30 kDa to ~105 kDa. RNA adapter ligation, IP-western, reverse transcription, DNA adapter ligation, and PCR amplification were performed in duplicates by Eclipse Bioinnovations Inc (San Diego, www.eclipsebio.com) according to the published single-end seCLIP protocol
GEO Accession
GSM4388159