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. Author manuscript; available in PMC: 2017 Dec 14.
Published in final edited form as: Chembiochem. 2016 Nov 11;17(24):2316–2323. doi: 10.1002/cbic.201600493

Figure 1.

Figure 1

A) Schematic representation of AlphaScreen assay format showing excitation of streptavidin-conjugated donor bead at 680 nm, production of singlet oxygen (1O2), and emission by anti-Flag-conjugated acceptor beads at 615 nm when a complex of biotinylated (bio) 4×PLXLE containing peptide and Flag-PHD2 is formed. ZF = zinc finger, PH = prolyl hydroxylase domain. B) AlphaScreen employing a biotinylated 4×PXLE peptide. Flag-PHD2 was omitted, EDTA (1 mM) was included, or Flag-PHD2 was substituted by Flag-PHD1 as indicated. n=3 per group. Error bars represent standard deviation. C) Inhibition of AlphaScreen signal from interaction of Flag-PHD2 (present in Sf9 extracts) and the biotinylated 4×PXLE peptide by non-biotinylated 4×PXLE (competitor) peptide with the identical amino acid sequence. n=3 per group. Error bars represent standard deviation. D) Pilot experiment for determining Z' parameter using competitor peptide. n=384. Error bars represent standard deviation.