EZH2(Y641F) TR-FRET Assay Kit
The EZH2 (Y641F) TR-FRET Assay Kit is designed to measure activity of the mutant EZH2 complex (EZH2 (Y641F)/EED/SUZ12/RbAp48/ AEBP) in a homogeneous 384 reaction format. This FRET-based assay requires no timeconsuming washing steps, making it especially suitable for high throughput screening applications. The EZH2 (Y641F) TR-FRET Assay Kit comes in a convenient format, with histone H3 peptide substrate, a Tb-labeled antibody against methylated K27 residue of Histone H3, S-adenosylmethionine, methyltransferase assay buffer, TR-FRET detection buffer, dye-labeled acceptor, and purified EZH2 (Y641F) complex for 384 enzyme reactions. The key to the EZH2 (Y641F) Assay Kit is a highly specific antibody that recognizes methylated Histone H3K27. 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 240 minutes. Next, antibody is added. Finally, dye-labeled acceptor is added followed by fluorescence reading.
| Catalog Number | Components | Amount | Storage | |
| 51017 | EZH2 (Y641F)/EED/SUZ12/RbAp48/AEBP2 | 2 x 50 µg | -80°C | Avoid freeze/ thaw cycles! |
| 52120 | 20 µM S-adenosylmethionine | 2 x 250 µl | -80°C | |
| 52089 | Tb-labeled antibody | 5 µl | -80°C | |
| Biotinylated histone H3 peptide substrate* | 500 µl | -80°C | ||
| 52170-A | 4x HMT Assay Buffer 2A | 3 ml | -20°C | |
| Dye-labeled acceptor | 2 x 10 µl | -20°C | ||
| TR-FRET Detection Buffer | 4 ml | -20°C | ||
| White, Nonbinding Corning, low volume, microtiter plate | 1 | Room Temp. | ||
1. Dillon, S.C., et al. Genome Biology 2005; 6:227.
2. Morin, R.D., et al. Nat Genet. 2010, 42(2):181.
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