GSM1561388: H3K27ac ChIP-seq, Prdm16 KO BAT; Mus musculus; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Histone
Antigen
H3K27ac
Cell type
Cell type Class
Adipocyte
Cell type
Brown adipocytes
NA
NA
Attributes by original data submitter
Sample
source_name
Prdm16 Knockout BAT
strain
Myf5Cre-Prdm16(fl/fl) C57Bl6
tissue
Brown adipose tissue
antibody
H3K27ac (Abcam ab4729)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Interscapular BAT was isolated and processed for each sacrificed mouse. The tissues were immediately minced, crosslinked in 1 % formaldehyde/PBS. MED1 ChIPs were performed using EGS as an additional crosslinker. For each fat pad, material from 3-4 mice was used for sequencing. To prepare ChIP extracts, crosslinked tissue was suspended in a ChIP buffer containing 20mM Tris, 0.1% SDS, 1% Triton-100, 150 mM NaCl, 1 mM EDTA, protease inhibitors. Samples were incubated at 4 C for 10 min, and subjected to microtip probe sonication under conditions optimized for IP efficiency. Extracts were cleared of insoluble lipid by microfuge centrifugation at full speed followed by extract transfer to a new tube. The process was repeated until minimal lipid accumulated at the top of the extract after centrifugation. ChIP-Seq libraries were produced and sequenced according to Illumina protocols. Libraries were multiplexed for sequencing.