Fluorescein-labeled methylated histone peptides (Karebay) were used for dissociation constant (Kd) determination by serial dilution in triplicate. The peptides used were H3K9me3 [NH2-ARTKQTARK(me3)STGG-minipeg-K(5-fam)-NH2] or H3K27me3 [Ac-QLATKAARK(me3)SAPA-minipeg-K(5-fam)-NH2] at a concentration of 1 nM. Serial dilutions were prepared in tris-buffered saline buffer [25 mM tris and 125 mM NaCl (pH 8)]. FP (fluorescence polarization) values were read on a DTX 880 Multimode Detector Beckman Coulter plate reader with excitation filter at 485 nm and two emission filters at 535 nm equipped with polarizers. Data were fit to a nonlinear regression equation using GraphPad Prism 4 software.

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