Anti-FITC Antibodies

Flow cytometry using anti-FITC antibody. Fixed and permeabilized A549 cells were labeled with mouse anti-tubulin antibody (1 µg/mL), followed by detection using a FITC-conjugated goat anti-mouse IgG secondary antibody (right peak, green). To confirm specificity, the cells were labeled with a mouse isotype control and stained using FITC goat anti-mouse IgG (middle peak, pink). Quenching of fluorescence upon pre-incubation of 1:100 of FITC Recombinant Polyclonal Antibody (Cat. No. 710083) is shown as middle peak, red. Unstained control cells are as shown (left peak, purple). The representative 10,000 cells were obtained and analyzed for each sample using Attune NxT Flow Cytometer.

Western blot analysis of fluorescein isothiocyanate (FITC) antibody detection. Analysis was performed by loading various amounts of a FITC-BSA conjugate per well onto a 4–20% Tris-glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% milk in TBST for at least 1 hour at room temperature. FITC-BSA was detected at ~72 kD using an FITC Monoclonal Antibody (#8) (Cat. No. MA5-14709) at a dilution of 1 µg/mL in 5% milk in TBST overnight at 4°C on a rocking platform, followed by a Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP (Cat. No. 31430) at a dilution of 1:50,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura Extended Duration Substrate (Cat. No.34076).