Fig. (2) Design of AlphaScreen-based 14-3-3 binding assays. (A) Biotin-14-3-3 proteins are coupled to streptavidin-coated donor beads (D) while GST-R18 is attached to anti-GST antibody-coated acceptor beads (A). (B) Biotinylated pS259-Raf-1 peptides are coupled to streptavidin-coated donor beads while GST-14-3-3 proteins are attached to acceptor beads with coated anti-GST antibodies. Interaction of 14-3-3 with R18 or pS259-Raf-1 leads to generation of luminescence signals at the indicated wavelength.