Suitable for studying enzyme kinetics and HTS applications.
Test Principle
The PRMT9 Homogeneous Assay Kit is designed to measure PRMT9 activity for screening and profiling applications. PRMT9 is a histone methyltransferase that exhibits methylation activity toward an arginine residue of SAP145. The PRMT9 Homogeneous Assay Kit comes in a convenient AlphaLISA format, with GST-tagged PRMT9 substrate, primary antibody, methylation assay buffer, and purified PRMT9 for 384 enzyme reactions. The key to the PRMT9 Homogeneous Assay 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 PRMT9 enzyme is incubated with the substrate. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts.
Contraindications
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.
Kit Components
PRMT9, 1x40ug 250uM S-adenosylmethionine, 1x400ul Primary antibody 28, 1x20ul PRMT9 substrate, 1x200ul 4x PRMT9 assay buffer, 3x1ml 4x Detection buffer, 1x2ml Protective film (to seal the wells)
Storage and Stability
Store powder at 4°C liquid at -20°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.
References
1. Li Y., et al., J. Neurosci. 2015; 35(37): 12890-12902.