Selected scFv antibodies were screened for the binding of TGEV-infected cells. For this, ST cells were grown to 60 to 70% confluence on coverslips and infected with TGEV at an MOI of 0.01 for 24 h. Cells were fixed in 4% paraformaldehyde and permeabilized with 0.5 % Triton X-100 for 10 min. The cells were then incubated with 100 μL of purified scFv antibody (20 μg/mL), and an unrelated scFv, ZW 88, was used as a negative-control antibody. The bound scFv antibodies were visualized using FITC-conjugated anti-His-tag antibody (Abcam) and a Nikon Eclipse 80i fluorescence microscope. The 293T cells transfected with p3 × FLAG-TGEV-S were also analyzed by IFA using scFv antibody as the primary antibody at 48 h post-transfection.
Fitc conjugated anti his tag antibody
Manufactured by Abcam
FITC-conjugated anti-His-tag antibody is a laboratory reagent used to detect and visualize proteins with a His-tag (polyhistidine tag) sequence. The antibody is conjugated with the fluorescent dye FITC (Fluorescein isothiocyanate), allowing for fluorescent detection of the His-tagged proteins.
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2 protocols using fitc conjugated anti his tag antibody
1
Screening scFv Antibodies for TGEV Binding
2
Verifying tCoa-NGR Binding to CD13/αvβ3 Receptors
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FACS analysis was employed to verify the binding of tCoa-NGR to CD13/αvβ3 integrin receptors, as previously described [14 (link), 45 (link), 46 (link)]. Briefly, human endothelial cells were mixed with either tCoa-NGR fusion proteins or tCoa (0.1 ng/106 cells in ice-cold FACS buffer) for 60 minutes at room temperature under light-protected condition. Finally, after addition of FITC-conjugated anti-His tag antibody (Abcam) signals for FITC was captured using BD FACS Calibur.
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