SET Domain-containing Protein 8 (SET8) is a methyltransferase that selectively mono-methylates histone H4 at lysine residue 20 (H4K20), an event proven to have an important role in chromatin structure and transcriptional activation. TheSET8 Methyltransferase Inhibitor Screening Assay provides a convenient method for screening human SET8 inhibitors. The transfer of the methyl group from SAM by SET8 (provided in the kit) to the acceptor peptide (H4K20) generates S-adenosylhomocysteine, which is rapidly converted to urate and hydrogen peroxide (H2O2) using an enzyme mixture provided in the kit. The H2O2 formed is quantified using 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) to produce the highly fluorescent compound resorufin (excitation 530-540 nm; emission 585-595 nm).
MT Assay Buffer: 1 vial/20 ml; -20°CMT Assay Buffer Additive: 1 vial/200 µl; -20°CMT Enzyme Mixture: 3 vials/300 µl; -80°CMT Fluorometric Mixture: 3 vials; -20°CMT Assay S-Adenosylmethionine: 3 vials; -80°C SET8 (human recombinant): 2 vials/100 µl; -80°CDMSO Assay Reagent: 1 vial/1 ml; Room temperature HCl Assay Reagent (20 mM): 1 vial/1 ml; -20°CSET8 Assay Acceptor Peptide: 2 vials/600 µl; -20°C MT Assay AdoHcy Positive Control: 1 vial/200 µl; -80°CHalf Volume 96-Well Solid Plate (black): 1 plate; Room temperature96-Well Cover Sheet: 1 cover Room; temperature