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Histone methyltransferase (HMT) activity/inhibition assay kit (H3-K9)
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Cat. No.:
OB0125WXX-047
Synonyms:
Histone methyltransferase (HMT); HMT activity/inhibition assay kit (H3-K9)
Size:
96T
Specification
Description:
Histone methyltransferase (HMT) activity/inhibition assay kit (H3-K9) is an epigenetic test to determine HMT activity/inhibition (H3-K9) using an innovative colorimetric assay that provides greater assay flexibility and is suitable for manual or high-throughput analysis.
Background:
Epigenetic inactivation of genes plays a crucial role in many important human diseases, especially cancer. A major mechanism of epigenetic inactivation of genes is the methylation of CpG islands in genomic DNA by DNA methyltransferases. HMTs control or regulate DNA methylation through chromatin-dependent transcriptional repression or activation. They transfer 1-3 methyl groups from S-adenosyl-l-methionine to lysine and arginine residues in histones. Inhibition of HMT leads to silencing of gene expression, and HMT inhibitors are currently being developed for a variety of therapeutic or experimental applications. SUV39-h1, SETDB1, etc., are HMTs that catalyze the methylation of histone H3 lysine 9 (H3-K9) in mammalian cells. H3-K9 methylation is mediated by the formation of HP3-K9 methylation. K9 methylation mediates heterochromatin formation by forming binding sites for HP1 and is also involved in silencing gene expression at non-chromatin sites. Assays for HMT activity/inhibition are necessary.
Assay Type:
Activity/inhibition assay
Assay Principle:
The histone substrate is stably captured in the wells of the reagent strip during the assay. The HMT enzyme transfers a methyl group from Adomet to the histone H3 substrate, methylating the substrate lysine 9. Methylated histone H3-K9 is recognized by high-affinity antibodies. The ratio or amount of methylated H3-K9 is proportional to enzyme activity and can be quantified by an HRP-conjugated secondary antibody chromogenic system. HMT activity is then calculated from the amount of methylated H3-K9 converted by HMTs.
Assay Target:
HMT
Target Species:
Universal
Research Area:
Methylation research; Obesity
Sample Type:
Biological samples
Kit Components:
HK1 (10X wash buffer); HK2 (Histone assay buffer); HK3 (Adomet); HK4 (Biotinylated substrate, 25 μg/mL); HK5 (HMT standard, 10 μg/mL); HK6 (Capture antibody, 100 μg/mL); HK7 (Detection antibody, 200 μg/mL); HK8 (Developing solution); HK9 (Stop solution); Control enzyme (300 μg/mL); 8 well assay strip (With frame)
Applications:
Histone methyltransferase (HMT) activity/inhibition assay kit (H3-K9) is ideally suited for the specialized determination of activity/inhibition of individual HMTs targeting lysine residues at different sites.
Storage:
Store at -20°C (HK3, HK4, HK5, HK7) and 4°C away from light (others).