GSM2790896: anti Brd4cachexia JQ1+; Mus musculus; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
Brd4
Cell type
Cell type Class
Muscle
Cell type
Skeletal muscle
NA
NA
Attributes by original data submitter
Sample
source_name
adult skeletal muscle
tissue
adult skeletal muscle
age
10 weeks
chip antibody
BRD6
strain
BALB/c
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Frozen tissues were pulverized in Covaris tissue TUBEs (Covaris 520001) using a chilled hammer on a cold metallic block, in dry ice. Powdered tissue was resuspended in PBS, cross-linked in 1% formaldehyde for 15 minutes. After blocking crosslink with 0.125M glycine for 5 minutes and washes with PBS plus protease and phosphatase inhibitors, pellet was resuspended in Farnham buffer (5 mM PIPES pH 8.0; 85 mM KCl; 0.5% NP-40) and briefly homogenized with a tissue homogenizer. Cells were subsequently lysed in RIPA buffer (1x PBS; 1% NP-40; 0.5% sodium deoxycholate; 0.1% SDS). 100mg of starting tissue was used for each antibody. Chromatin was sonicated to fragments length of approximately 0.5Kb and immunoprecipitated with 3.5µg of BRD4 antibody. Immunoprecipitation procedures were performed as in Proserpio et al., 2013. For ChIPSeq, 10ng of immuno-precipitated DNA fragments were used to prepare ChIP-seq libraries with Ovation SP Ultralow DR Multiplex system (Nugen) following the manufacturer’s protocol.