Specifications

  • Assay Type Activity Assay
  • Sample Types Lysates, Purified Enzyme Systems
  • Assay Duration 30 Minutes
  • Samples/Plate 40 in Duplicate
  • Readout Fluorescent, 510 nm emission / 450 nm excitation
  • Standard Curve
  • Description

    Assay Principle: The Histone Demethylase (HDM) Fluorescent Activity Kit allows activity measurement of any Histone Demethylase system by measuring formaldehyde. Run the demethylase reaction, add the Formaldehyde Detection Reagent and read the fluorescent signal generated. Lab-to-lab comparisons are possible with inclusion of a formaldehyde reference standard. No separation or washing is required. The kit has been validated for both LSD1 and JMJD2A Histone Demethylases (HDMs). Please read the complete kit insert before performing this assay

    Background: Histone demethylases (HDMs) catalyze the site-specific demethylation of methyl-lysine residues in histones to dynamically regulate chromatin structure, gene expression, and potentially other genomic functions. At present, there are two known classes of HDMs: the flavin adenine nucleotide (FAD)-dependent Lysine Specific Demethylase 1 (LSD1) family and the Fe(II)-dependent Jumonji C (JmjC) family. Although they employ different cofactors and catalytic mechanisms, both classes produce formaldehyde as a byproduct of the demethylation reaction. HDMs have proven difficult to quantitatively assay owing to their relatively low turnover numbers. For this reason, understanding of HDMs kinetic properties, substrate specificities, and reaction mechanisms is limited.