Figure 5
From: Identification of a GTP-bound Rho specific scFv molecular sensor by phage display selection
Binding activity of scFv C1 compared to scFv C1-N1N2. (A) scFv C1 and (B) scFv C1-N1N2 crude extracts analyzed by (1) Western Blot and (2) ELISA. (1) scFvs were resolved on 12,5% SDS-PAGE, immunoblotted with c-myc antibody, and visualized by enhanced chemiluminescence as described in Methods. (2) scFvs from crude extract were analysed for binding to GST-RhoA and GST-RhoAQ63L protein immobilized on an ELISA plate. Bound scFvs were detected with horseradish peroxydase-labeled anti-c-myc using TMB as substrate. Results are expressed as absorbance at 480 nm. Graphs are representative of 6 experiments, each performed in duplicate.