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Applications: Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Background: Histone H4 (R3) methyltransferases are enzymes thatcatalyze the transfer of a methyl group from the cofactor S-adenosylmethionine toarginine 3 residue of histone H4. This group of enzymes includes PRMT1, PRMT3, and PRMT5.
Contraindications: Avoid green and blue dyes that absorb light in the Alphascreen® signal emission range (520-620 nm), 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.
Description: The Histone H4(R3) Universal Methyltransferase Assay Kit is designed for the detection of histone H4(R3) methyltransferase (HMT) activity in screening and profiling applications. The keyto the Histone H4(R3) Universal Methyltransferase Assay Kit is a highly specificantibody that recognizes methylated R3 residue of Histone H4. With this kit, only threesimple steps on a microtiter plate are required for methyltransferase detection. First, S-adenosylmethionine is incubated with a sample containing assay buffer andmethyltransferase enzyme for one hour. Next, primary antibody is added. Finally, theplates are treated with an HRP-labeled secondary antibody followed by addition of theHRP substrate to produce chemiluminescence that can then be measured using achemiluminescence reader.
Storage Stability: 12 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Supplied As: Kit comes in a convenient format, with a 96-well plate precoated with histone H4 peptide substrate, purified PRMT1, PRMT3 and PRMT5 enzymes, and reagents for 100 enzyme reactions.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Dillon S.C., et al. 2005. Genome Biology 6:227.
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