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Applications: Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Background: JMJD2E, also known as KDM4DL (lysine (K)-specific demethylase 4D-like) is a JmjC-domain protein that exhibits demethylation activity toward H3-K9Me3 and H3-K36Me3.
Contraindications: Avoid 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 JMJD2E Homogeneous Assay Kit is designed to measure activity of JMJD2E for screening and profiling applications, using AlphaLISA® technology. The key to the JMJD2E Homogeneous Assay Kit is a highly specificantibody that recognizes demethylated substrate. With this kit, only three simple steps on amicrotiter plate are required for methyltransferase detection. First, a sample containingJMJD2E enzyme is incubated with the biotinylated substrate for two hours. Next, acceptorbeads and primary antibody are added, then donor beads, followed by reading the Alphacounts.
Storage Stability: 12 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Supplied As: Kit comes in a convenient AlphaLISA® format, with a 384-well plate, biotinylated histone H3 peptide substrate, primary antibody, demethylase assay buffer, and purified JMJD2E for 384 enzyme reactions.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Whetstine, J.R. et al., Cell 2006; 125: 467.
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