| Assay Method Information | |
| | In Vitro Methyltransferase Activity Assay |
| Description: | Recombinant PRC2 (EZH2-Y641F) was purchased from Active motif company, S-adenosyl-methionine (SAM) and Poly-L-lysine (PLL) were purchased from Sigma-Aldrich company, and H3(1-50)K27me1 peptide was purchased from Cisbio company. The detection system uses LANCEUltra system (Perkinelmer company). In the enzyme activity test, compounds to be tested were diluted by 8 gradient points in a ratio of 1:3, and added into a reaction plate, then 100 ng recombinase was added. And then a buffer [20 mM Tris pH8.5, 2 mM MgCl2, 0.01% Tween-20, 1 mM TCEP] containing 2.5 μM SAM/250 nM H3 (1-50)K27me1 premixtures was added, thus the enzyme reaction was started at room temperature. After reacting for 3 hours, a test solution premixed with PLL, detection antibody and Ulight was added, and reacted for 1h at room temperature, then the fluorescence value was read on a Tecan infinite pro. |
| Affinity data for this assay | |
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