PRMT8 Homogeneous BioAssay™ Kit (Protein Arginine N-methyltransferase 8, Histone-arginine N-methyltransferase 8, HRMT1L3, HRMT1L4)

RMT8 is a histone methyltransferase that exhibits methylation activity toward histones H2A and H4.

Intended Use:
Suitable for use in studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.

Test Principle:
The PRMT8 Homogeneous BioAssay™ Kit is designed to measure PRMT8 activity for screening and profiling applications. PRMT8 is a histone methyltransferase that exhibits methylation activity toward histones H2A and H4. The PRMT8 Homogeneous BioAssay™ Kit comes in a convenient AlphaLISA format, with a 384-well plate, biotinylated histone H4 peptide substrate, primary antibody, methylation assay buffer, and purified PRMT8 for 384 enzyme reactions. The key to the PRMT8 Homogeneous BioAssay™ Kit is a highly specific antibody that recognizes methylated histone substrate. With this kit, only three simple steps on a microtiter plate are required for methyltransferase detection. First, a sample containing PRMT8 enzyme is incubated with the biotinylated substrate for one hour. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts.

Contraindications:
Avoid 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:
PRMT8, 1x50ug
100uM S-adenosylmethionine, 1x250ul
Primary antibody 4, 1x400ul
Biotinylated histone H4 peptide substrate, 1x400ul
4x HMT Assay Buffer 2A, 1x3ml
4x Detection Buffer, 1x2ml

Storage and Stability:
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.