PRMT5 is a histone methyltransferase that exhibits methylation activity toward H4-R3. PRMT5 is a critical regulatory-protein linked to genome organization,cell cycle regulation,and stem cell differentiation. PRMT5 transfers methyl groups from S-adenosylmethoinine (SAM) to form mono and symmetric di-methylated arginine residues in histone proteins,transcription factors,and other regulatory proteins. Given its regulatory function,PRMT5 is believed to contribute to tumorigenesis. Its dysregulated activity has been associated with many cancers including lymphomas,lung,and breast cancer. Additionally,PRMT5 has been linked to other diseases including kidney and heart disease as well as neurological disorders such as Huntington’s and Alzheimer’s diseases.Intended Use:Suitable for screening small molecular inhibitors for drug discovery and HTS applications.Test Principle:The PRMT5 TR-FRET BioAssay Kit is designed to measure PRMT5 activity in a homogeneous 384 reaction format. PRMT5 is a histone methyltransferase that exhibits methylation activity toward H4-R3. This FRET-based assay requires no time-consuming washing steps,making it especially suitable for high throughput screening applications. The PRMT5 TR-FRET Assay Kit comes in a convenient format,with histone H4 peptide substrate,a Eu-labeled antibody,S-adenosylmethionine,methyltransferase assay buffer,FRET detection buffer,dye-labeled acceptor,and purified PRMT5 for 384 enzyme reactions. The key to the PRMT5 TR-FRET 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,S-adenosylmethionine is incubated with a sample containing assay buffer and methyltransferase enzyme for 120 minutes. Next,antibody is added. Finally,dye-labeled acceptor is added followed by fluorescence detection.Kit Components:PRMT5/MEP50,1x40ug60uM S-adenosylmethionine,1x250ulEu-labeled antibody,1x5ulBiotinylated histone H4 peptide substrate,1000 rxns4x PRMT5 assay buffer,3x1ml,add 10ul of 0.5M DTT/vial before use.Dye-labeled acceptor,1x20ulTR-FRET Detection Buffer,1x4mlWhite,Nonbinding Corning,low volume,microtiter plate,1x plateStorage 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.