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Histone methyltransferase (HMT) activity/inhibition assay kit (H3-K27)
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Cat. No.:
OB0125WXX-048
Synonyms:
Histone methyltransferase (HMT); HMT activity/inhibition assay kit (H3-K27)
Size:
96T
Specification
Description:
Histone methyltransferase (HMT) activity/inhibition assay kit (H3-K27) is an epigenetic test kit for the detection of HMT activity/inhibition (H3-K27) by a colorimetric assay that can be analyzed manually or in high throughput and is flexible.
Background:
The epigenetic inactivation of genes is crucial 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 of histones. Inhibition of HMT leads to the silencing of gene expression, and HMT inhibitors are being developed for various therapeutic or experimental applications. G9a and polymerase group enzymes are HMTs that catalyze the methylation of histone H3 at lysine 27 (H3-K27) in mammalian cells. H3-K27 methylation mediates the formation of heterochromatin through the formation of polymerase binding sites and is also involved in the silencing of nonchromatin. H3-K27 methylation mediates heterochromatin formation through the formation of polymerase binding sites and is also involved in silencing gene expression at non-chromatin sites. It has also been found that increased global H3-K27 methylation is associated with pathological processes. Assays for HMT activity/inhibition are necessary.
Assay Type:
Activity/inhibition assay
Assay Principle:
During the assay, the histone substrate is stably captured on the wells by a biotin-streptavidin conjugate, and the HMT enzyme transfers a methyl group from Adomet to the histone H3 substrate, methylating the substrate lysine 4. Methylated histone H3-K27 is recognized by high-affinity antibodies. The ratio or amount of methylated H3-K27 is proportional to enzyme activity and can be quantified by an HRP-conjugated secondary antibody chromogenic system. HMT activity is then calculated based on the amount of methylated H3-K27 converted by HMTs.
Assay Target:
HMT
Target Species:
Universal
Research Area:
Methylation research; Obesity
Sample Type:
Biological samples
Kit Components:
HE1 (10X wash buffer); HE2 (Histone assay buffer); HE3 (Adomet); HE4 (Biotinylated substrate, 25 µg/ml); HE5 (HMT standard, 10 µg/ml); HE6 (Capture antibody, 100 µg/ml); HE7 (Detection antibody, 100 µg/ml); HE8 (Developing solution); HE9 (Stop solution); Control enzyme (150 µg/ml); 8 well assay strip (With frame)
Applications:
Histone methyltransferase (HMT) activity/inhibition assay kit (H3-K27) is ideally suited to specifically assay the activity/inhibition of individual HMTs targeting lysine residues at different sites.
Storage:
Store at -20°C (HE3, HE4, HE5, HE7, control enzyme) and 4°C away from light (others).