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Applications: Great for studying enzyme kinetics and HTS applications.
Contraindications: Green and blue dyes that absorb light in the AlphaScreen signal emission range (520-620 nm), such as Trypan Blue. Avoid the use of the potent singlet oxygen quenchers such as sodium azide (NaN3) or metal ions (Fe2+, Fe3+, Cu2+, Zn2+ and Ni2+). The presence of >1% RPMI 1640 culture medium leads to a signal reduction due to the presence of excess biotin and iron in this medium. MEM, which lacks these components, does not affect AlphaScreen assays.
Description: The JMJD2D Homogeneous Assay Kit is designed to measure JMJD2D activity for screening and profiling applications. JMJD2D, also known as KDM4D, is a JumonjiC (JmjC) domain containing histone lysine demethylase that exhibits demethylation activity toward H3-K9Me3 and H3-K36Me3. The key to the JMJD2D Homogeneous Assay Kit is a highly specific antibody that recognizes demethylated substrate. With this kit, only three simple steps on a microtiter plate are required for methyltransferase detection. First, a sample containing JMJD2D enzyme is incubated with the biotinylated substrate. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts. Our histone demethylase assays utilize highly specific antibodies that recognize demethylated products. First, a sample containing the enzyme is incubated with a biotinylated substrate. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts, as shown above.
Storage Stability: At least one year from date of receipt when stored as directed.
Supplied As: The JMJD2D Homogeneous Assay Kit comes in a convenient AlphaLISA® format (Scheme 1), with biotinylated histone H3 peptide substrate, primary antibody, demethylase assay buffer, and purified JMJD2D for 384 enzyme reactions.
Warnings: This product is for research purposes only and not for human or therapeutic use. This product should be considered hazardous. Do not ingest, inhale, get in eyes, on skin, or on clothing. If so, wash thoroughly.
Biosafety Level: Not applicable (BSL-1)
References: 1. Whetstine, J.R., et al. Cell 2006; 125(3): 467-481.
2. Fodor, B.D., et al. Genes & Dev. 2006. 20: 1557-1562.
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