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H3k9m2

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H3K9m2 is a histone modification antibody that specifically recognizes dimethylation at lysine 9 of histone H3. This histone modification is associated with transcriptional repression and heterochromatin formation.

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Lab products found in correlation

Pcr purification kit, by Thermo Fisher Scientific (2 mentions) Ace h3, by Merck Group (2 mentions) H3k4m3, by Merck Group (2 mentions)

2 protocols using h3k9m2

1

Chromatin Profiling of Muscle Cell Epigenetics

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Quantitative PCR (qPCR), immunoblotting, and chromatin immunoprecipitation (ChIP) assay were performed as previously described (15 (link)). For the ChIP assay, C2C12 or primary human myotubes were fixed with 1% formaldehyde and sonicated to produce chromatin lysates. The lysates were precleared with Protein-G agarose beads and immunoprecipitated overnight with antibodies against Ace-H3 (Millipore), Ace-H4 (Millipore), H3K4m3 (Millipore), H3K9m2 (Abcam), or control IgG in the presence of BSA and salmon sperm DNA. The next day, Protein-G agarose beads were added to each immunoprecipitation reaction for 1 h, followed by an extensive wash and reverse crosslink. DNA was eluted from the beads and purified using a PCR Purification kit (Invitrogen) and subsequently analyzed by qPCR using primers located on the proximal Baf60c and Deptor promoters. Primers are available upon request.
Meng Z.X., Wang L., Xiao Y, & Lin J.D. (2014). The Baf60c/Deptor Pathway Links Skeletal Muscle Inflammation to Glucose Homeostasis in Obesity. Diabetes, 63(5), 1533-1545.
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2

ChIP Assay for Histone Modifications

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ChIP assay was performed according to the protocol developed by Upstate Biotechnology as described (Meng et al., 2013 (link)). Briefly, treated myotubes were fixed with 1% formaldehyde and sonicated to produce chromatin lysates. The lysates were pre-cleared with Protein-G agarose beads and immunoprecipitated overnight with antibodies against Ace-H3 (Millipore, 06–599), H3K4m3 (Millipore, 07–473), H3K9m2 (Abcam, ab1220), or control IgG in the presence of BSA and salmon sperm DNA. The next day, Protein-G agarose beads were added to each immunoprecipitation reaction for 1 h, followed by extensively wash and reverse crosslink. DNA was eluted from the beads and purified using a PCR Purification Kit (Invitrogen) and subsequently analyzed by qPCR using primers located on the proximal promoter regions of mouse Baf60c and Deptor genes (Table S1).
Meng Z.X., Gong J., Chen Z., Sun J., Xiao Y., Wang L., Li Y., Liu J., Shawn Xu X.Z, & Lin J.D. (2017). Glucose sensing by skeletal myocytes couples nutrient signaling to systemic homeostasis. Molecular cell, 66(3), 332-344.e4.
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