Suitable for studying enzyme kinetics and HTS applications.

The PRMT5 Homogeneous BioAssay™ Kit is designed to measure PRMT5 activity for screening and profiling applications. PRMT5 is a histone methyltransferase that exhibits methylation activity toward H4-R3. The PRMT5 Homogeneous BioAssay™ Kit comes in a convenient AlphaLISA format, with biotinylated histone H4 peptide substrate, primary antibody, methylation assay buffer, and purified PRMT5 for 384 enzyme reactions. The key to the PRMT5 Homogeneous BioAssay™ Kit is a highly specific antibody that recognizes methylated substrate. With this kit, only three simple steps on a microtiter plate are required for methyltransferase detection. First, a sample containing PRMT5 enzyme is incubated with the biotinylated substrate for two hours. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts.

Green and blue dyes that absorb light in the AlphaScreen signal emission range (520-620nm), 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.

PRMT5/MEP50, 2x40ug 60uM S-adenosylmethionine, 1x250ul Primary antibody 4-3, 1x100ul Biotinylated histone H4 peptide substrate, 500 rxns 4x PRMT5 assay buffer (add DTT before experiment), 3x1ml 4x Detection buffer, 1x2ml

Store powder at 4°C liquid at -20°C/-70°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.