ChIP-Atlas: SRX2787198

Antigen

Cell type Class: Blood
Cell type: Macrophages
MeSH Description: The relatively long-lived phagocytic cell of mammalian tissues that are derived from blood MONOCYTES. Main types are PERITONEAL MACROPHAGES; ALVEOLAR MACROPHAGES; HISTIOCYTES; KUPFFER CELLS of the liver; and OSTEOCLASTS. They may further differentiate within chronic inflammatory lesions to EPITHELIOID CELLS or may fuse to form FOREIGN BODY GIANT CELLS or LANGHANS GIANT CELLS. (from The Dictionary of Cell Biology, Lackie and Dow, 3rd ed.)

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: DNA was extracted using the PureGene DNA extraction kit (Gentra Systems). Samples from BCGvaccinated and non-vaccinated, Mtb-infected and non-infected BMDMs were crosslinked with 37% w/v formaldehyde for 20 minutes at RT and immediately quenched for 5 minutes with 1M Glycine at RT. The formaldehyde fixed samples were then sonicated to 150500 bp using an E220 sonicator (Covaris), and subsequently ChIP-DNA was prepared using a manual chromatin immunoprecipitation method with AntibodiesAntigen incubation for 18 hours, followed by bead incubation for 150 minutes, and 7 5minute washing steps. Approximately 1x106 cells were used for each ChIP and ~50,000 cells for the input. The following antibodies were used: H3K4me1 (Company: CST, Cat. No.: 5326P, Lot No.: 1), H3K4me3 (Thermo Fisher Scientific, MA5-11199), H3K27ac (Abcam, Ab4729, Lot No.: GR2005631). ChIP and Input libraries were prepared using the MicroPlex Library Preparation Kit v2 (C05010013, Diagenode), with alterations including: PCR enrichment (9 to 18 cycles) prior to size selection, and use of Ampure beads for size selection (250-350 bp).