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Encompassing Amino Acids: 2-751
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 10 µl reaction mix containing assay buffer with 20 mM HEPES (pH 7.4), 50 mM NaCl 500 µM α-ketoglutarate, 25 µM iron, 2mM ascorbic acid, 0.01% Tween-20, 0.5 µM biotinylated peptide substrate, and JARID1B (20 - 100 ng) added to the wells. Add antibody against demethylated K4 peptide. Incubate for 30 min, then Streptavidin-conjugated secondary antibody followed by Alpha Screening detection.
Background: Histone demethylase that demethylates Lys-4 of histone H3, thereby playing a central role in histone code. Does not demethylate histone H3 Lys-9 or H3 Lys-27. Demethylates trimethylated, dimethylated and monomethylated H3 Lys-4. Acts as a transcriptional corepressor for FOXG1B and PAX9. Favors the proliferation of breast cancer cells by repressing tumor suppressor genes such as BRCA1 and HOXA5. In contrast, may act as a tumor suppressor for melanoma.
Description: Human JARID1B, also known as PLU-1 and KDM5B (GenBank Accession No. NM_006618), a.a. 2-751 with N-terminal His-FLAG-tag, MW=88 kDa, expressed in Sf9 cells via a baculovirus expression system.
Format: Aqueous buffer solution
Formulation: 40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 0.04% Tween-20, 80 µg/mL FLAG peptide and 20% glycerol.
Genbank: NM_006618
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal His-FLAG-tags
Uniprot: Q9UGL1
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 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.
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