Human JARID1B, also known as KDM5B and PLU-1 (GenBank Accession No. NM_006618), a.a. 2- 1544(end) with N-terminal FLAG-Avi-tag, MW=179 kDa, expressed in Sf9 cells via a baculovirus expression system.

Q9UGL1

JARID1B, PLU-1, KDM5B, Retinoblastoma-Binding Protein 2 Homolog 1, RBP2-H1, CT31, Cancer/Testis Antigen 31, PLU-1, Jumonji, AT Rich Interactive Domain 1B, Lysine-Specific Demethylase 5B

0.11 pmol/min/µg

10 µl reaction mix containing 20 mM HEPES (pH 7.4), 50 mM NaCl, 500 µM α-ketoglutarate, 25 µM iron, 2 mM ascorbic acid, 0.01% Tween-20, 1 mM TCEP, 0.5 µM biotinylated peptide substrate, and JARID1B. Incubate 1 hour at RT with slow shaking. Add antibody against demethylated product and anti-mouse acceptor beads. After 30 minutes, add 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% Tween-20, 20% glycerol, 80 μg/ml FLAG peptide, and 0.2 mM TCEP

At least 6 months at –80°C.

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

1. Yamane K, et al. (2007). Mol. Cell 25 (6): 801–12.
2. Barrett A, et al. (2007). Int. J. Cancer 121 (2): 265–75.

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

Demethylases

Enzymes