Epi-C offers standardised and custom epigenetic assays for
HDAC1, HDAC4, HDAC6, SIRT1, P300 and JMJ.
Histone deacetylase (HDAC) enzymes act by catalytially removing acetyl groups from histone lysine residues.
Deactylation of specific histone residues (such as H3K9-14Ac) results in chromatin condensation and transcriptional repression.
The assay involves a double-step reaction and the product is acquired via fluorescence.
The JmjC family comprises 30 members that share a JmjC domain. To date, 18 have been shown to possess demethylase activity toward H3K4, H3K9, H3K27, H3K36 and H4K20. Although all members react with α-ketoglutarate (α-KG) in a Fe (II) ion-dependent manner, not all have been shown to be catalytically active.
In addition, others domains may mediate demethylase activity independently of the JmjC domain. Specific substrate assays are performed via fluorescent reader based on JMJ sub-family member.
SIRT1, a Class III HDAC, is a beta-NAD-dependent deacetylase enzyme.
P53K374-389 acetylation is the substrate for SIRT1 activity. Reaction is due to the cleavage of Beta-NAD to nicotinamide (NAM) and ADP-Ribose.
NAM cleavage produces free ammonia, which is detected by fluorescence reader using a developer solution.