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. Author manuscript; available in PMC: 2018 Feb 5.
Published in final edited form as: J Mol Biol. 2016 Nov 16;429(4):562–573. doi: 10.1016/j.jmb.2016.11.008

Fig. 8.

Fig. 8

ELISA assay indicates that RasIn2 has higher affinity for H-Ras(G12V)-GppNHp than Raf1-RBD. (a) Schematic of the ELISA assay. Biotinylated H-Ras(G12V) was captured on the ELISA plate via streptavidin–biotin link. FLAG-tagged RasIn2 or Raf-RBD was then incubated with the ELISA plate, followed by an anti-FLAG antibody, and then a secondary antibody conjugated to horseradish peroxidase (HRP). The plate was then washed and incubated with tetramethylbenzidine (TMB). (b) The OD450 of the ELISA readout plotted over different concentrations of ligand (RasIn2 or Raf-RBD). RasIn2 shows ~10-fold higher specific signal level than Raf-RBD.