The EZH2 Homogeneous Assay Kit is designed to measure EZH2 activity for screening and profiling applications. EZH2 is a histone methyltransferase that exhibits methylation activity toward H3-K27. The key to the EZH2 Homogeneous Assay Kit is a highly specific antibody that recognizes methylated substrate. With this kit, only three simple steps on a microtiter plate are required for methyltransferase detection. First, a sample containing EZH2 enzyme is incubated with the histone substrate for one hour. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts.

AlphaLISA anti-rIgG acceptor beads, 5 mg/ml (PerkinElmer #AL104C)
AlphaScreen Nickel donor beads, 5 mg/ml (PerkinElmer #AS101D)
Optiplate -384 (PerkinElmer #6007290)
AlphaScreen microplate reader

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.