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. 2009 Nov 6;38(2):e11. doi: 10.1093/nar/gkp899

Figure 5.

Figure 5.

The AlphaScreen assay is adaptable to measuring binding of proteins harboring other chromatin-associated domains. The double tudor domain-containing JMJD2A protein (100 nM) interacts with b-H3K4me3 (white), and b-H4K20me3 (black) peptides (A). His6-tagged JMJD2A protein was titrated with 10–1000 nM b-H3K4me3 peptide (B). Unlabeled H3K4me3 peptide competed with 100 nM b-H3K4me3 for binding to 100 nM His-JMJD2A (C).The PHD-containing RAG2 protein was similarly titrated with 10–1000 nM b-H3K4me3 peptide (D). Unlabeled H3K4me3 peptide competed with b-H3K4me3 (50 nM) for binding to 5 nM GST-RAG2 (E). Data are shown as the mean and standard deviation of two or four replicate measurements for matrix titrations (A, C) and peptide competition assays (B and D), respectively.