Human JMJD3 (KDM6B) histone demethylase (a.a. 1043-end), Genbank Accession No. NM_001080424, with C-terminal FLAG-tag, MW=71 kDa, expressed in Sf9 cells via a baculovirus expression system.

O15054

JMJD3, KDM6B

1.1 pmole/min/μg

10 µl reaction mix containing assay buffer with 20 mM HEPES (pH 7.4), 150 mM NaCl 500 µM α-ketoglutarate, 25 µM iron, 2 mM ascorbic acid, 0.01% Tween-20, 0.5 µM biotinylated peptide substrate, and JMJD3 (100 – 500 ng) added to the wells. Add antibody against demethylated K27 peptide. Incubate for 30 min, then Streptavidin-conjugated secondary antibody followed by Alpha Screening detection.

40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl and 20% Glycerol.

Aqueous buffer solution

>6 months at –80°C.

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

1. Y. Xiang, et al., Cell Research (2007) 17:850–8572.
2. S. Hong, et al. Proc. Natl. Acad. Sci. USA (2007) 104(47):18439-44.
3. Swigut T, Wysocka J. Cell (2007) 131(1):29-3.

Application References:
1. A small molecule modulates Jumonji histone demethylase activity and selectively inhibits cancer growth (2013)
2. Identification of small molecule inhibitors of Jumonji AT-rich interactive domain 1B (JARID1B) histone demethylase by a sensitive high throughput screen (2013)

Avoid freeze/thaw cycles.

Demethylase

Recombinant Protein