DESCRIPTION

• Notes: The buffer in this kit was reformulated in May of 2015 with a different reducing agent to improve assay performance. The old formulation can still be purchased upon special request.
• Shiptemp: -80°C (dry ice)
• Synonyms: JHDM3B, KDM4B
• Warnings: Avoid freeze/thaw cycles.
• Category: Demethylase/Assay Kit
• Background: JMJD2B is a JmjC-domain protein that exhibits demethylation activity toward H3-K9Me3 and H3-K36Me3.
• References: 1. Whetstine JR et al. (2006) Cell 125: 467. 2. Fodor, B.D., Genes & Dev. 2006. 20: 1557-1562. APPLICATION REFERENCE:Nakagawa-Yagi, Y., et al. BMC Complement Altern Med. 12(1):101 (2012).
• Description: The JMJD2B Homogeneous Assay Kit is designed to measure activity of the JMJD2B for screening and profiling applications, using AlphaLISA® technology. The key to the JMJD2B Homogeneous Assay Kit is a highly specific antibody that recognizes demethylated substrate. With this kit, only three simple steps on a microtiter plate are required for methyltransferase detection. First, a sample containing JMJD2B enzyme is incubated with the biotinylated substrate. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts.
• Methodology: AlphaLISA®
• Supplied As: The JMJD2B Homogeneous Assay Kit comes in a convenient format, with a 384-well plate, biotinylated histone H3 peptide substrate, primary antibody, demethylase assay buffer, and purified JMJD2B for 384 enzyme reactions.
• Unspsc Code: 41121811
• Unspsc Name: Laboratory microchemistry kits
• Applications: Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
• Product Type: Assay Kit
• Biosafety Level: Not applicable (BSL-1)
• Contraindications: Avoid green and blue dyes that absorb light in the Alphascreen® signal emission range (520-620 nm), such as Trypan Blue. Avoid the use of the potent singlet oxygen quenchers such as sodium azide (NaN3) or metal ions (Fe2+, Fe3+, Cu2+, Zn2+ and Ni2+). The presence of >1% RPMI 1640 culture medium leads to a signal reduction due to the presence of excess biotin and iron in this medium. MEM, which lacks these components, does not affect Alphascreen® assays.
• Storage Stability: 12 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
• Scientific Category: Demethylase
• Instructions for Use: See assay kit datasheet for detailed protocol.