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 key to the Histone H4(R3) Universal Methyltransferase 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 for one hour. Next, primary antibody is added. Finally, the plates are 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.

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.

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.

COMPONENTS

See assay kit data sheet for detailed protocol.

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.

Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.

Dillon S.C., et al. 2005. Genome Biology 6:227.

Avoid freeze/thaw cycles.

Methyltransferase

Assay Kit