Next, they added a solution of captopril with various concentrations to phosphate buffer of gold nanoparticles - fluorescein isothiocyanate
, and they studied the fluorescent spectrum of its emitted fluorescein isothiocyanate
5 for fluorescein isothiocyanate
, which covers most food products' pH.
5% Tween 20 in PBS) for 5 min each, and then exposed to goat anti-mouse IgG conjugated with fluorescein isothiocyanate
(FITC) (1:100) (BOSTER, China).
After centrifugation at 400g for 5 min, we suspended the leukocytes in 490 [micro]L binding buffer (Annexin V-FITC Apoptosis Detection Kit, Biovision) and incubated the leukocyte suspension with 5 [micro]L each of reagent set-supplied, undiluted fluorescein isothiocyanate
-labeled annexin V (BioVision) and reagent set-supplied, undiluted propidium iodide (BioVision) at room temperature for 5 min in the dark.
After washing, these cells were first incubated with a monoclonal anti-insulin antibody (1:200 dilution; Sigma, Madrid, Spain) for 2 hr and then for 1 hr with a secondary anti-body, anti-mouse IgG-conjugated fluorescein isothiocyanate
(IgG-FITC; 1:200 dilution; Sigma), both at room temperature.
The fluorometer is used to measure the fluorescence signal of a suspension of microspheres with immobilized fluorescein isothiocyanate
specific antibody CRY104 labeled with fluorescein isothiocyanate
(Biotech Frontiers, North Ryde, Australia) for enumeration.
Secondary antibodies, all obtained from SigmaAldrich, were as follows: fluorescein isothiocyanate
(FITC)-conjugated anti-mouse IgG ([gamma]-chain specific), peroxidase-conjugated anti-mouse IgG ([gamma]-chain specific), FITC-conjugated anti-rabbit IgG (whole molecule); and peroxidase-conjugated anti-rabbit IgG (whole molecule).