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 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 JMJD2E enzyme is incubated with the biotinylated substrate for two hours. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alphacounts.

JMJD2E, KDM4DL, lysine (K)-specific demethylase 4D-like, AlphaLISA, Alphascreen, Alpha Lisa, Alpha screen

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.

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.

See assay kit data sheet for detailed protocol.

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.

Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.

Whetstine, J.R. et al., Cell 2006; 125: 467.

AlphaScreen^® and AlphaLISA^® are registered trademarks of PerkinElmer, Inc.

Avoid freeze/thaw cycles.

Demethylase

Assay Kit