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. 2015 Nov 2;5:15916. doi: 10.1038/srep15916

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Binding of KRAS4b-FME to GST-RAF-RBD is detected using the alpha assay. The nickel chelate donor bead recognizes the His6 tag on Nanodiscs (containing 30% DMPS) and transmits a singlet oxygen to the glutathione sepharose acceptor bead that recognizes the GST tag on the RAF-RBD. This alpha signal is observed only with GMP-PNP bound KRAS4b-FME and not non-processed KRAS (a). The alpha signal is dependent on the concentration of GST-RAF-RBD and is not observed with GDP bound KRAS-FME (b).