GSM1280296: NELF ChIP-Seq in HeLa; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
TFs and others
Cell type Class
Attributes by original data submitter
NELF-A (A-20) (Santa Cruz, sc-23599)
Sequenced DNA Library
HeLa cells were grown to 0.5 million cells/ml and crosslinked with 1% formaldehyde in media for 10 minutes before glycine addition to 125 mM. After washing and lysis, cells were sonicated in RIPA buffer (50 mM Tris, pH 7.6; 150 mM NaCl; 1 mM EDTA; 0.25% sodium deoxycholate; 1% IGEPAL CA-630) on ice using a Fisher Model 550 Sonic Dismembrator. For each ChIP, sonicated supernatant from 5 million cells was incubated at 4°C overnight with 10 µg of Pol II (Santa Cruz, sc-899), NELF (sc-23599), or DSIF (sc-28678) antibody, and then 1 hr with 50 µl Protein G-Sepharose bead slurry (Sigma, P3296). Beads were washed in columns with 15 ml ice cold RIPA buffer and rinsed with 10 ml ice cold PBS. Immunocomplexes were eluted, incubated at 65°C overnight to reverse crosslinks, and treated with RNase A and Proteinase K. DNA was isolated by MinElute PCR purification kit (QIAGEN, 28004). Library preparation was performed by the University of Iowa DNA Facility using the Ovation SP Ultralow DR Multiplex System (Nugen, 8033-32) for Mondrian.