Intended Use:Suitable for studying enzyme kinetics and HTS applications.
Test Principle:The PRMT8 Chemiluminescent BioAssay™ Kit is designed to measure PRMT8 activity for screening and profiling applications. The PRMT8 Chemiluminescent BioAssay™ Kit comes in a convenient format with a 96-well plate precoated with histone H4 peptide substrate, the antibody against methylated arginine 3 (R3) residue of Histone H4, the secondary HRP-labeled antibody, S-adenosylmethionine, methyltransferase assay buffer, and purified PRMT8 enzyme for 100 enzyme reactions. The key to the PRMT8 Chemiluminescent BioAssay™ 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:PRMT8 His-FLAG-tags, 1x10ug20uM S-adenosylmethionine*, 1x250ulPrimary antibody 4, 1x100ulSecondary HRP-labeled antibody 2, 1x10ul4x HMT assay buffer 2, 1x3mlBlocking buffer, 1x50mlHRP chemiluminescent substrate A (transparent bottle), 1x6mlHRP chemiluminescent substrate B (brown bottle), 1x6ml96-well plate precoated with histone substrate, 1x plate* Decreasing the S-adenosylmethionine concentration will make the assay more sensitive to test inhibitors.
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.
