GSM5928547: IP Stimulated RNase p65 1; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
RELA
Cell type
Cell type Class
Blood
Cell type
Jurkat
Primary Tissue
Blood
Tissue Diagnosis
Leukemia Acute Lymphocytic
Attributes by original data submitter
Sample
source_name
Jurkat cells
treatment
PMA + Ionomycin stimulation + Rnase treatment
antibody
anti-p65 rabbit polyclonal (Invitrogen; 51-0500)
Sequenced DNA Library
library_name
GSM5928547
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were cross-linked with 1% parafolmaldehyde for 10 minutes and glycine was added to quench the reaction. The cell lysates were prepared as described (Varshney et al., 2018). The chromatin was sonicated in Bioruptor (Diagenode) at high power for 40 min (30 s on/off). 40 micrograms of the sonicated chromatin were used for each IP. 5% of each extract used in the IPs were taken as inputs.For p65, 7.5ug of anti-p65 rabbit polyclonal (Invitrogen; 51-0500) or Rabbit IgG Isotype Control (ThermoFisher; 02-6102) antibody were added to samples and incubated overnight at 4oC, before incubation with 30ul Protein G Dynabeads (Life Technologies) for an additional 2h. Beads were eluted as desribed by Varshney et al before Proteinase K treatment and reverse crosslinking at 42oC overnight. Eluted DNA was purified by QIAquick PCR purification kit (QIAGEN). Libraries were prepared using NEBNext® Ultra™ II DNA Library Prep Kit and NEBNext® Multiplex Oligos (Index Primers Set 1) for Illumina.