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Applications: Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Background: JARID1C, also known as SMCX (Smcy homolog, X-linked), KDM5C, or XE169, is a JumonjiC (JmjC) and ARID domain-containing histone lysine demethylase that exhibits demethylation activity toward di- and trimethyl-lysine 4 (H3K4me2/3) on histone H3.
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.
Description: The JARID1C Homogeneous Assay Kit is designed to measure activity of the JARID1C for screening and profiling applications, using AlphaLISA® technology. The key to the JARID1C 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 demethylase activity detection. First, a sample containing JARID1C enzyme is incubated with the biotinylated substrate. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts.
Storage Stability: At least 6 months at -80°C.
Supplied As: The JARID1C Homogeneous Assay Kit comes in a convenient AlphaLISA® format, with biotinylated histone H3 peptide substrate, primary antibody, demethylase assay buffer, and purified JARID1C for 384 enzyme reactions.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Iwase S, Lan F, Bayliss P, et al. Cell 2007; 128(6):1077-1088
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