The EpiQuik™ DNMT3B Activity/Inhibitor Screening Assay Core Kit is a convenient set of tools that allows the experimenter to selectively screen DNMT3B activity/inhibitors in an ELISA-like format. The selective inhibition of DNMT3B may lead to demethylation and expression of silenced tumor suppressor genes. Thus, the selective DNMT3B inhibitors could be a new addition to cancer therapeutic agents. The kit is ready-to-use and provides all the essential components needed to carry out a successful DNMT3B activity/inhibitor screening experiment without the need for radioactivity or any special equipment. The kit has the following advantages:
Note: This "core" kit does not come with enzymes included.
Principle & ProcedureThe EpiQuik™ DNMT3B Activity/Inhibitor Screening Assay Core Kit is designed for screening Dnmt 3B inhibitors. In an assay with this kit, the unique cytosine-rich DNA substrate is stably coated on the strip wells. These wells are specifically treated to have a high DNA absorption ability. A DNMT3B (not included) enzyme transfers a methyl group to cytosine from Adomet to methylate DNA substrate. The methylated DNA can be recognized with an anti-5-methylcytosine antibody. The ratio or amount of methylated DNA, which is proportional to enzyme activity, can then be colorimetrically quantified through an ELISA-like reaction.
Fig. 1. Schematic procedure for using the EpiQuik DNMT3B Activity/Inhibitor Screening Assay Core Kit.
Fig. 2. Activity of recombinant DNMT3B at different concentrations was measured using the EpiQuik™ DNMT3B Activity/Inhibitor Screening Assay Core Kit.
Fig. 3. Recombinant DNMT3B was incubated with substrate and inhibitor BD-11. DNMT3B activity was measured in the presence or absence of inhibitors.
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