The PRMT5 Homogeneous Assay Kit is designed to measure PRMT5 activity for screening and profiling applications. PRMT5 is a histone methyltransferase that methylates histone H4-R3. The Assay Kit relies on a highly specific antibody that recognizes the methylated form of the substrate. With this kit, only three simple steps on a microtiter plate are required for methyltransferase detection. First, a sample containing PRMT5 enzyme is incubated with the biotinylated substrate for two hours. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts.

Illustration of the assay principle: The PRMT5/MEP50 enzyme methylates the biotinylated histone H4 peptide substrate. The methylated form of the peptide is recognized by a highly specific rabbit antibody, which binds to anti-Rabbit acceptor beads, while the biotin binds to streptavidin-conjugated donor beads, bringing the acceptor and donor beads in close proximity. Upon excitation of the donor bead a singlet oxygen is generated by the donor bead, which excites the acceptor bead and emits light proportionally to the level of interaction. AlphaLISA™ immunoassays are a no-wash alternative to ELISA immunoassays. These assays are robust and ideal for a minimal hands-on approach.

*NOTE: As of April 2023, this kit has been re-optimized for performance.

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