Three types of arginine methylation exist in mammalian cells: monomethylarginine (MMA), asymmetric dimethylarginine (ADMA) and symmetric dimethylarginine (SDMA). Type I PRMTs ((CARM1, PRMT1, PRMT2, PRMT3, PRMT6, and PRMT8) catalyze ADMA, while type II PRMTs (e.g. PRMTs 5 and 7) catalyze SDMA. Type II PRMTs are found to be strongly implicated in diseases like cancer. For example, PRMT5 plays a role in the repression of certain tumor suppressor genes such as RB tumor suppressors while PRMT7 overexpression is observed in breast cancer.
Source:Recombinant protein corresponding to aa2–end from human PRMT5/MEP50, fused to N-terminal FLAG-tag (PRMT5) and N-terminal His-tag (MEP50), co-expressed in Baculovirus infected Sf9 cell expression system.
Molecular Weight:~73kD (PRMT5), 37.5kD (MEP50)
Assay Conditions:50ul reaction mix (20mM phosphate buffer pH 7.4, 20uM S-adenosyl methionine, and 10-100ng methyltransferase PRMT5) add to the wells coated with the substrate. Incubate at room temperature for 1 hr. Add antibody against methylated R3 residue of histone H4, incubate 1 hr. Add secondary HRP-labeled antibody and incubate 30 min. Finally, add HRP chemiluminescent substrates and read luminesence.
Storage and Stability:Aliquot to avoid repeated freezing and thawing and store at -70°C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
