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Fig. 1 | BMC Research Notes

Fig. 1

From: Validation of extracellular ligand–receptor interactions by Flow-TriCEPS

Fig. 1

Development of Flow-TriCEPS as a method to determine ligand binding on living cells. a Flow cytometric analysis to determine binding of transferrin coupled to TriCEPS (TRFE-TriCEPS) to the surface of living MDA-MB-231 cells (red line). Competition assay by adding increasing amount of unlabeled TRFE. A representative experiment out of two is shown. b Addition of BSA, cannot compete the TRFE-TriCEPS-PE signal on MDA-MD-231 cells (red line). c Flow cytometric analysis to determine binding of antiCD71-PE labelled on MDA-MB-231 cells transfected with siRNA TFR1 (Hs_TFR1_5) at different time points, Igg-PE was used as control. d Flow cytometric analysis to determine binding of TRFE-TriCEPS on MDA-MB-231 cells transfected with siRNA TFR1 (Hs_TFR1_5) at different time points, cells labelled with PE-Streptavidin were used as control. Similar results were obtained by using Hs_TFR1_11 and Hs_TFR1_7 siRNA for 72 h. A representative experiment out of 3 is shown. e Confocal microscopy using TriCEPS-TAMRA to detect binding of TRFE-TriCEPS-TAMRA on the surface of living HEK293 cells. As control of the specificity TRFE-TriCEPS was out-competed with excess of unlabeled TRFE

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