The FBXL10 (KDM2B) Homogeneous Assay Kit is designed to measure the activity of the FBXL10 for screening and profiling applications. FBXL10, also known as KDM2B, JHDM1B, and CXXC2, is a histone lysine demethylase that exhibits demethylation activity toward trimethyl-lysine 4 (H3K4me3) and dimethyl-lysine 36 (H3K36me2) on histone H3. The key to the FBXL10 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 demethylase activity detection. First, a sample containing FBXL10 enzyme is incubated with the biotinylated substrate. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts.

AlphaLISA anti-rabbit IgG acceptor beads5 mg/ml (PerkinElmer #AL104L)

AlphaScreen Streptavidin-conjugated donor beads 5 mg/ml (PerkinElmer #6760002S)

Optiplate - 96 (PerkinElmer #6005290)

AlphaScreen microplate reader

Adjustable micropipettor and sterile tips

KDM2B, JHDM1B, CXXC2

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 culture medium RPMI 1640 at >1% leads to 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 protocol.

At least one year from date of receipt when stored as directed.

Great for studying enzyme kinetics and HTS applications.

Iwase, S., Lan, F., Bayliss, P., et al. Cell 2007; 128(6):1077-1088.

Avoid multiple freeze/thaw cycles

Demethylase

Assay Kit