H3K4me3_M1Salt_3
Mus musculus
Protocols: Primary bone-marrow-derived murine macrophages were grown in RPMI1640 (GIBCO) supplemented with 10% FCS, 10mM HEPES, 1% Penicillin-Streptomycin, 5µM beta-mercaptoethanol at a density of 40 million cells per 150cm(2) cell culture flask. After attachment for 2h cells were stimulated with 10ng/ml LPS (Sigma-Aldrich) [M(LPS)] for 24h. 40mM NaCl were added where applicable. 40 million macrophages per group were used to isolate nuclei by centrifugation through a dense sucrose cushion. Subsequently, samples were digested with micrococcal nuclease (Sigma-Aldrich) to generate native chromatin templates consisting mainly of mononucleosomes. Abundance of mononucleosomal DNA was confirmed by gel electrophoresis. Chromatin was extracted with RIPA buffer and cellular debris removed by centrifugation. Subsequently, chromatin was precipitated over night at 4°C with anti-H3K4me3 (NEB #9751 S) bound to Dynabeads® Protein A (Invitrogen #10002D). MinElute PCR Purification Kit (QIAGEN) was used to purify immunoprecipitated DNA after Proteinase K (Sigma-Aldrich) digestion at 65°C for 4h. 50ng of purified DNA was used for library preparation with the TruSeq ChIP sample preparation kit (Illumina) according to the manufacturer's instructions.
ENA-FIRST-PUBLIC
2015-02-26T17:03:04Z
ENA-LAST-UPDATE
2018-03-08T23:43:40Z
External Id
SAMEA3267657
INSDC center name
MDC
INSDC first public
2015-02-26T17:03:04Z
INSDC last update
2018-03-08T23:43:40Z
INSDC status
public
Submitter Id
E-MTAB-3343:H3K4me3_M1Salt_3
broker name
ArrayExpress
cell type
macrophage
common name
house mouse
sample name
E-MTAB-3343:H3K4me3_M1Salt_3
scientific_name
Mus musculus
sex
male
stimulus
10ng/ml LPS 24h
strain
C57BL/6JOlaHsd