Human UTX, also known as KDM6A (GenBank Accession No. NM_021140) (amino-acids 1-1401) with N-terminal FLAG-tag, MW = 155 kDa, expressed in Sf9 cells via a baculovirus expression system.

O15550

UTX, KDM6A

0.096 pmol/min/μg

10 µl reaction mix containing assay buffer with 50 mM HEPES (pH 7.4), 1mM TCEP, 100 µM alpha-ketoglutarate, 25 µM iron, 0.1mM ascorbic acid, 0.02% Tween-20, 0.5 µM biotinylated peptide substrate, and UTX (10 – 50ng) added to the wells. Add antibody against di-methylated 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, 0.04% Tween20, 20% glycerol, 80 μg/ml flag peptide and 0.2mM TCEP

At least 6 months at –80°C. Avoid freeze/thaw cycles. Storing diluted enzyme is not recommended, if necessary.

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

1. Greenfield A, et al. (1998). Hum. Mol. Genet. 7 (4): 737–42.
2. Agger K, et al. (2007). Nature 449 (7163): 731–4.

Application Reference(s):
1. Inhibition of KDM4A activity as a strategy to suppress interleukin-6 production and attenuate colitis induction (2017)
2. Identification of the KDM2/7 histone lysine demethylase subfamily inhibitor and its antiproliferative activity (2013)

Avoid freeze/thaw cycles. Storing diluted enzyme is not recommended, if necessary, use carrier protein (BSA 0.1 – 0.5%).

Demethylase

Recombinant Protein