GSM2771531: HUDEP2 BCL11A-ER-V5 ChIP-seq Input 1; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Input control
Antigen
Input control
Cell type
Cell type Class
Blood
Cell type
HUDEP-2
NA
NA
Attributes by original data submitter
Sample
source_name
HUDEP2 BCL11A-ER-V5 ChIP-seq Input 1
treatment
Induction of the BCL11A-ER-V5 system was achieved with the addition of 4-hydroxytamoxifen (dissolved in 100% ethanol) at a final concentration of 10-7 M (0.1 nmol). Cells were harvested after 24 hours of induction for downstream experimentation.
cell line
HUDEP-2
antibody
IgG (sc-2028)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ChIP experiments were performed as previously described (Schmidt D, Wilson MD, Spyrou C, Brown GD, Hadfield J, Odom DT. ChIP-seq: using highthroughput sequencing to discover protein-DNA interactions. Methods. Jul 2009;48(3):240-248). Briefly, approximately 5x107-1x108 cells were used for each IP. Cells were crosslinked with 1 % formaldehyde for 10-15 min at RT and the reaction was quenched with glycine at a final concentration of 125 mM. Crosslinked cells were then lysed and sonicated to obtain ~200–300 bp fragments of chromatin. Crosslinked DNA was pulled down at 4 °C overnight using antibody against V5-tag (R960CUS, Life Technologies), ZBTB7A (Santa Cruz Biotechnology Inc, sc-33683 x and eBioscience, 14-3309-82) or control anti-goat IgG (sc-2028). Chromatin was then reverse crosslinked and eluted at 65°C overnight and DNA was purified.If ChIP material was subjected to sequencing, DNA was purified using a Qiagen MinElute Reaction Clean-up kit. Standard Illumina TruSeq