Universal Histone Demethylase (HDM) Fluorescent Activity kit allows all known HDMs to be measured in low, medium and high throughput fashion. Run the demethylase reaction and after completion simply add the supplied Formaldehyde Detection Reagent to each well and read the fluorescent signal generated. Excitation at 450nm/Emission at 510nm using an adjustable gain plate fluorimeter.
Formaldehyde is a common byproduct formed in oxidative demethylation. Examples of formaldehyde-producing enzymes include histone demethylases (HDMs) that modify methylated histones. Lysine-specific HDMs were first discovered in 2004 and are currently among the most actively studied formaldehyde-producing enzymes. 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 the LSD1 and JmjC HDMs employ different cofactors and catalytic mechanisms, both produce formaldehyde as a byproduct of the demethylation reaction. Despite their biological importance, HDMs have proven difficult to quantitatively assay owing to their relatively low turnover numbers, hindering our understanding of their kinetic properties, substrate specificities, and reaction mechanisms.
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