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Fig. 2 | BMC Research Notes

Fig. 2

From: Production of a mono-biotinylated EGFR nanobody in the E. coli periplasm using the pET22b vector

Fig. 2

Mono-biotinylated EL2BH binds to EGFR in EGFR expressing cells. a Immuno-precipitation and western blotting of EGFR from A431 cell lysates using EL2BH nanobody, or commercial poly-biotinylated EGFR antibody as positive control, both pre-conjugated to streptavidin agarose beads. Streptavidin agarose beads in PBS with or without antibody were used as a negative control. b Flow cytometry analysis of A431 cells stained with EL2BH nanobody pre-coupled to streptavidin-Alexa Fluor 488, compared to positive control antibody pre-coupled to streptavidin-Alexa Fluor 488 and vehicle control with streptavidin-alexa 488 only. c Immunostaining of EGFR-negative and EGFR-overexpressing HEK293T cells (EGFR OV) with EL2BH pre-coupled to Streptavidin Alexa Fluor 488 (green), counterstained with Hoechst 3342 (blue). d Flow cytometry of EGFR-negative and EGFR-overexpressing HEK293T cells (EGFR OV) with EL2BH pre-coupled to streptavidin-Alexa Fluor 488. The flow cytometry data were analyzed using Flowjo software

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