HMTs

HMTs

Histone peptide substrate labelled with proprietary long lifetime 9-aminoacridine (9AA) fluorophore and incorporating an aromatic moiety which reduces the FLT of 9AA

  • PKMT catalysed methylation of the histone substrate prevents cleavage by the protease, Endo-LysC, thereby maintaining modulation of the FLT of 9AA
  • Hence, PKMT activity is reported by a decrease in the measured FLT of the assay

 

 

G9a Assay – Methylation of H3K9

  • The FLEXYTE™ G9a assay employs a tailored histone H3 peptide substrate to efficiently report methylation at Lys9
  • The assay was validated for inhibition studies using the known BIX-01294 inhibitor whose measured IC50 was consistent with the literature value
  • The assay was shown to be compatible with a wide range of SAM concentrations and suitable for HTS applications (Z’-factor > 0.7)

Set 7/9 Assay – Methylation of H3K4

  • The FLEXYTE™ Set 7/9 assay employs a tailored histone H3 peptide substrate to efficiently report methylation at Lys4
  • The assay was validated for inhibition studies using the generic SAM competitive inhibitor, S-adenosylhomocysteine (SAH)
  • The assay was shown to be compatible with a wide range of SAM concentrations and suitable for HTS applications (Z’-factor > 0.8)

Protein Arginine Methyltransferases (PRMT)

PRMT Assay Principle

  • Histone peptide substrate labelled with proprietary long lifetime 9-aminoacridine (9AA) fluorophore and incorporating an aromatic moiety which reduces the FLT of 9AA
  • PRMT catalysed methylation of the histone substrate prevents cleavage by the protease, Endo-ArgC, thereby maintaining modulation of the FLT of 9AA
  • Hence, PRMT activity is reported by a decrease in the measured FLT of the assay

PRMT5 Assay – Methylation of H4R3

FLEXYTE™ methyltransferase assays have recently been extended to PRMTs, such as human PRMT5 which symmetrically methylates H4R3

The assay employs a tailored H4 peptide substrate whose FLT is reduced until exposed to the protease, Endo-ArgC. Methylation of Arg3 by PRMT5 confers protease protection, leading to a concentration and time dependent decrease in the FLT of the peptide reporter.

For more information, please see the contact us page.

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