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Applications: Great for studying enzyme kinetics and HTS applications.
Contraindications: Green and blue dyes that absorb light in the AlphaScreen signalemission range (520-620 nm), such as Trypan Blue. Avoid the use of the potent singlet oxygenquenchers such as sodium azide (NaN3) or metal ions (Fe2+, Fe3+, Cu2+, Zn2+ and Ni2+). Thepresence of culture medium RPMI 1640 at >1% leads to signal reduction due to the presence ofexcess biotin and iron in this medium. MEM, which lacks these components, does not affectAlphaScreen assays.
Description: The JMJD1A Homogeneous Assay Kit is designed to measure activity of the JMJD1A forscreening and profiling applications. JMJD1A, also known as KDM3A, is a JumonjiC (JmjC)domain-containing histone lysine demethylase that exhibits demethylation activity toward monoanddimethyl-lysine 9 on histone H3. The key to theJMJD1A Homogeneous Assay Kit is a highly specific antibody that recognizes demethylatedsubstrate. With this kit, only three simple steps on a microtiter plate are required for demethylaseactivity detection. First, a sample containing JMJD1A enzyme is incubated with the biotinylatedsubstrate. Next, acceptor beads and primary antibody are added, then donor beads, followed byreading the Alpha-counts.
Storage Stability: Stable at least 6 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.
Supplied As: The JMJD1A Homogeneous Assay Kit comes in a convenient AlphaLISA® format, with biotinylated histone H3 peptide substrate, primary antibody, demethylase assay buffer, and purified JMJD1A for 384 enzyme reactions.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Horton, J.R., et al. Nature Struct. & Mol. Biol. 2010; 17(1): 38-43.