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. Author manuscript; available in PMC: 2013 Feb 1.
Published in final edited form as: Anal Bioanal Chem. 2012 Jan 5;402(5):1889–1898. doi: 10.1007/s00216-011-5604-0

Figure 2. Schematic of assay principles.

Figure 2

(a) In the HTRF assay, the MBNL1-His6 and Biot-(CUG)12 complex is captured by the HTRF reagents anti-His-Terbium and streptavidin-XL665. The formation of protein-RNA complex brings terbium cryptate within close proximity to XL665 to allow time-resolved fluorescence energy transfer. (b) In the AlphaScreen assay, streptavidin-conjugated donor beads are brought into close proximity to nickel-coated acceptor beads upon capture by the protein-RNA complex. Excitation with 680nm light induces release from donor bead of singlet oxygen that is capable of energy transfer with thioxene derivatives in the acceptor bead, culminating in 520-620nm photoemission by acceptor bead.