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Applications: Great for studying enzyme kinetics and HTS applications.
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 PHF8 Homogeneous Assay Kit is designed to measure PHF8activity for screening and profiling applications. PHF8, also known as JHDM1F and KDM7B, is aJumonjiC (JmjC) domain containing histone lysine demethylase that exhibits demethylationactivity toward H3-K9Me2. The PHF8 Homogeneous Assay Kit comes in a convenientAlphaLISA® format (Scheme 1), with biotinylated histone H3 peptide substrate, primaryantibody, demethylase assay buffer, and purified PHF8 for 96 enzyme reactions. The key to thePHF8 Homogeneous Assay Kit is a highly specific antibody that recognizes demethylatedsubstrate. With this kit, only three simple steps on a microtiter plate are required formethyltransferase detection. First, a sample containing PHF8 enzyme is incubated with thebiotinylated 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 PHF8 Homogeneous Assay Kit comes in a convenient AlphaLISA® format (Scheme 1), with biotinylated histone H3 peptide substrate, primary antibody, demethylase assay buffer, and purified PHF8 for 96 or 384 enzyme reactions.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Horton, J.R., et al. Nature Structural & Molecular Biology 2010; 17(1): 38-43.
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