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Applications: Great for studying enzyme kinetics and HTS applications.
Background: UTX, also known as KDM6A, is a JmjC-domain protein that exhibits demethylation activity toward di- and trimethyl-lysine 27 (H3K27me2/3) on histone H3
Contraindications: 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 culture medium RPMI 1640 at >1% leads to 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 UTX Homogeneous Assay Kit is designed to measure the activity of UTX for screening and profiling applications. The key to the UTX 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, UTX 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 one year from date of receipt when stored as directed.
Supplied As: The UTX Homogeneous Assay Kit comes in a convenient AlphaLISA® format, with biotinylated histone H3 peptide substrate, primary antibody, demethylase assay buffer, and purified UTX for 384 enzyme reactions.
Uniprot: O15550
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1.Greenfield A, et al. 1998. Hum. Mol. Genet. 7(4): 737-42.
2. Agger K, et al. 2007. Nature 449(7163): 731-4.