Human JMJD2A, also known as JHDM3A and KDM4A (GenBank Accession No. NM_014663) (a.a. 1-350) with N-terminal His-tag, MW=42 kDa, expressed in an E. coli expression system.

O75164

JMJD2A, JHDM3A, KDM4A

0.084 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 JMJD2A (100 – 400 ng) added to the wells. Add antibody against demethylated K9 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, 250 mM imidazole, and 20% glycerol

Aqueous buffer solution

>6 months at –80°C.

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

1. Zhang, D., et al. (2005). Mol. Cell Biol. 25(15): 6404-6414.
2. Gray SG, et al. (2005). J. Biol. Chem. 280 (31): 28507–18.

Application Reference(s):
A scintillation proximity assay for histone demethylases (2014)

Avoid freeze/thaw cycles.

Demethylase

Recombinant Protein