organism: Mus musculus
description: Protocols: Mice were treated acutely with an intraperitoneal injection of dexamethasone or vehicle, and liver tissue harvested one hour later. Chromatin was prepared from flash-frozen liver tissue using the Active Motif ChIP-IT High Sensitivity kit (Active Motif). 25μg of chromatin was incubated overnight at 4C with a GR antibody cocktail (ProteinTech 24050-1-A (lot 00044414) and Cell Signaling D8H2 (lot 2)). To permit direct comparison between samples, and to control for technical variation between ChIP reactions, a spike-in ChIP normalisation strategy was employed, with 30ng spike-in chromatin (53083, Active Motif) and 2μg spike-in antibody (61686, Active Motif (lot 34216004)) being included in each IP reaction. To obtain sufficient DNA for next-generation sequencing, three IP reactions were carried out for each sample, and then pooled for the pull-down step. Antibody was pulled down using washed magnetic protein G agarose beads (ReSyn Biosciences). Beads were washed five times with AM1 Wash Buffer (Active Motif) then DNA eluted as per ChIP-IT kit instructions. ChIP-seq DNA was purified with the MinElute PCR Purification kit (Qiagen). Illumina TruSeq ChIP Library Preparation Kit, according to the manufacturer's protocol
attributes.tag: ENA first public
attributes.value: 2021-04-10
attributes.tag: ENA last update
attributes.value: 2021-03-16
attributes.tag: External Id
attributes.value: SAMEA8261953
attributes.tag: INSDC center alias
attributes.value: University of Manchester
attributes.tag: INSDC center name
attributes.value: University of Manchester
attributes.tag: INSDC first public
attributes.value: 2021-04-10T00:31:41Z
attributes.tag: INSDC last update
attributes.value: 2021-03-16T14:39:58Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-10224:WT_GRChIP-seq_input
attributes.tag: age
attributes.value: 10 to 12
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: organism part
attributes.value: liver
attributes.tag: sample name
attributes.value: E-MTAB-10224:WT_GRChIP-seq_input
attributes.tag: sex
attributes.value: male
attributes.tag: strain
attributes.value: C57BL/6