The cell surface reactivity of C6.5-diab and deB-C6.5-diab against HER2 positive SK-BR-3 tumor cells was determined by flow cytometry using a FACS Calibur (BD Biosciences, Mississauga, ON, Canada). Briefly, 2 × 105 cells were incubated with the indicated amounts of C6.5-diab or deB-C6.5-diab for 2 h on ice. After washing with 2% FBS in PBS, bound C6.5 diabody and deB-C6.5-diab were detected with an anti-His antibody (GE Healthcare) at a 1/800 dilution followed by a fluorescein isothiocyanate (FITC) labeled goat anti-mouse antibody (Sigma) at a 1/200 dilution. Cells were analyzed on a FACS Calibur following propidium iodide (Molecular Probes, Eugene, OR, USA) staining and cell surface reactivity determined for live cells.
Fitc labeled goat anti mouse antibody
Manufactured by Merck Group
Sourced in United States
The FITC-labeled goat anti-mouse antibody is a secondary antibody that binds to primary mouse antibodies. It is conjugated with the fluorescent dye FITC (Fluorescein Isothiocyanate), which allows for the detection and visualization of target proteins or cells in various applications, such as immunofluorescence or flow cytometry.
Automatically generated - may contain errors
2 protocols using fitc labeled goat anti mouse antibody
1
Flow Cytometric Evaluation of Tumor Cell Binding
2
Quantifying IL-1β in β-Glucan-Treated Zebrafish Cells
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To detect the presence of IL-1β in response to cells exposure to β-glucan, ZF4 cells monolayers, grown in 96-well plates at 28 ºC, were treated with β-glucan (10µg/mL) for 24h.
Then the cell monolayers were fixed for 15 minutes with a 4% paraformaldehyde solution followed by 15 minutes in cold methanol. For IL-1β detection, fixed ZF4 cells were incubated overnight with a mouse polyclonal antibody against rainbow trout IL1B (Nombela, 2017) kindly donated by Dr. Luis Mercado (Universidad Católica de Valparaiso, Chile) diluted 100-fold in
PBS with 0.3% Triton X-100 at 4ºC. The anti-IL1B antibody has been tested against ZF4 cells samples by ELISA (Fig. S3). After primary antibody incubation the cells were washed with PBS and incubated for 45 min with FITC labeled goat anti-mouse antibody (Sigma, USA) diluted 1:300 with 0.3% triton x100. Finally, the cells were then washed 3 times with PBS. To visualize the nuclei of the cells, the monolayers were counterstained with 4´-6-diamidino-phenylin-indole (DAPI) for 10 min. The cells were observed and photographed with an IN Cell Analyzer 6000 image system (GE Healthcare).
Then the cell monolayers were fixed for 15 minutes with a 4% paraformaldehyde solution followed by 15 minutes in cold methanol. For IL-1β detection, fixed ZF4 cells were incubated overnight with a mouse polyclonal antibody against rainbow trout IL1B (Nombela, 2017) kindly donated by Dr. Luis Mercado (Universidad Católica de Valparaiso, Chile) diluted 100-fold in
PBS with 0.3% Triton X-100 at 4ºC. The anti-IL1B antibody has been tested against ZF4 cells samples by ELISA (Fig. S3). After primary antibody incubation the cells were washed with PBS and incubated for 45 min with FITC labeled goat anti-mouse antibody (Sigma, USA) diluted 1:300 with 0.3% triton x100. Finally, the cells were then washed 3 times with PBS. To visualize the nuclei of the cells, the monolayers were counterstained with 4´-6-diamidino-phenylin-indole (DAPI) for 10 min. The cells were observed and photographed with an IN Cell Analyzer 6000 image system (GE Healthcare).
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