KDM5/JARID Activity Quantification Assay Kit (Fluorometric) (ab113464)

Ce kit est pour mesurer l’activité de KDM5 qui est une enzyme qui enlève les methyl des protéines et surtout des histones.

KDM5 a effectivement comme substrats, H3K4me3 et H3K4me2 et produit H3K4 mono-méthylée. Ce kit utilise donc un anticorps qui va reconnaitre H3K4 deméthylée/mono-méthylée.

Le standard est démethylé mais ceci ne veux pas dire non-méthylé dans ce contexte mais mono-méthylé.

Thank you for your interest in ab113455 & ab113464.

You are correct that both these kits measure demethylase activity. The major difference between these 2 kits is the substrate captured on the well. For ab113455, the substrate is a dimethylated histone H3-K4 substrate while the substrate used in ab113464 is a trimethylated histone H3-K4 substrate.

Due to these different substrates, ab113455 will detect LSD1 type demethylase activity which can remove mono and dimethyl groups while ab113464 will detect JARID type demethylase activity which removes trimethyl groups from H3 K4.

So it seems ab113464 would be the proper choice for your interest in studying H3K4me3 demethylation activity.

As far as we know neither of these kits have been used in published works.

I hope this is helpful. Please contact us again if you have any further questions.

Thank you for your call last week and for your patience while I have been in touch with my colleagues in the lab regarding ab113464.

The substrate is a short peptide, and the detection antibody binds to mono-methylated histone.

I hope this information will be useful, but if there is anything else that we can do for you please let me know and we'll be happy to help.

Thank you for contacting us. Tri-methylated histone H3-K4 is a substrate for JARID enzyme which removes methyl group from substrate, which is then detected by antibodies present in the kit. So the kit antibodies will detect the JARID de-methylated products. The amount of demethylated products is proportional to the enzyme activity of JARID which can be fluorometrically measured by reading the fluorescence in a fluorescent microplate reader at 530 excitation and 590 emissions. The whole procedure is explained in the protocol section of this kit; which can be downloaded by clicking the following link; https://www.abcam.com/ps/products/113/ab113464/documents/ab113464%20EpiSeeker%20KDM5%20JARID%20Activity%20Quantification%20Assay%20Kit-Fluorometric%20(Website).pdf I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.