PRMT5 is a histone methyltransferase that exhibits methylation activity toward H4-R3.
Intended Use:Suitable for studying enzyme kinetics and HTS applications.
Test Principle:The PRMT5 Chemiluminescent BioAssay™ Kit is designed to measure PRMT5 activity for screening and profiling applications. The PRMT5 Chemiluminescent BioAssay™ Kit comes in a convenient format, with a 384-well plate precoated with histone H4 peptide substrate, the antibody against methylated arginine3 residue of Histone H4, the secondary HRP-labeled antibody, S-adenosylmethionine, methyltransferase assay buffer, and purified PRMT5 enzyme for 384 enzyme reactions. The key to the PRMT5 Chemiluminescent Assay Kit is a highly specific antibody that recognizes methylated R3 residue of Histone H4. With this kit, only three simple steps on a microtiter plate are required for methyltransferase detection. First, S-adenosylmethionine is incubated with a sample containing assay buffer and methyltransferase enzyme. Next, primary antibody is added. Finally, the plate is treated with an HRP-labeled secondary antibody followed by addition of the HRP substrate to produce chemiluminescence that can then be measured using a chemiluminescence reader.
Contraindications:DMSO >1%, strong acids or bases, ionic detergents, high salt
Kit Components:PRMT5/MEP50 human enzyme, 1x40ug100uM S-adenosylmethionine, 1x250ulPrimary antibody 4-3, 1x100ulSecondary HRP-labeled antibody 2, 2x10ul4x PRMT5 assay buffer (add DTT before experiment), 2x3mlBlocking buffer, 2x50mlHRP chemiluminescent substrate A (transparent bottle), 2x6mlHRP chemiluminescent substrate B (brown bottle), 2x6ml384-well plate precoated with histone substrate, 1x plate
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.
