MUC13 expressing HPAF-II, AsPC-1 and Panc-1 cells were incubated with Tetramethylrhodamine (TRITC) labeled anti-MUC13 MAb (25 μg/ml) for 1 h at 4 °C and plated for 2 h at 37 °C for adherence. Cells were fixed and permeabilized with Triton X-100. Cells were then incubated with anti-HER2 antibody (Dako) or α-tubulin (negative control) for 1 h followed by incubation with FITC goat anti-Rabbit secondary antibody (catalog number 65-6111; Thermo Fisher Scientific) followed by confocal microscopy 22 (link).
Fitc goat anti rabbit secondary antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The FITC goat anti-Rabbit secondary antibody is a fluorescently labeled antibody that specifically binds to and detects rabbit primary antibodies. It is designed for use in various immunoassay techniques, such as flow cytometry, immunohistochemistry, and Western blotting, to visualize and analyze target proteins or cells of interest.
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Lab products found in correlation
4 protocols using fitc goat anti rabbit secondary antibody
1
Visualizing MUC13 and HER2 Protein Localization
2
Visualizing MUC13 and HER2 Protein Localization
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MUC13 expressing HPAF-II, AsPC-1 and Panc-1 cells were incubated with Tetramethylrhodamine (TRITC) labeled anti-MUC13 MAb (25 μg/ml) for 1 h at 4 °C and plated for 2 h at 37 °C for adherence. Cells were fixed and permeabilized with Triton X-100. Cells were then incubated with anti-HER2 antibody (Dako) or α-tubulin (negative control) for 1 h followed by incubation with FITC goat anti-Rabbit secondary antibody (catalog number 65-6111; Thermo Fisher Scientific) followed by confocal microscopy 22 (link).
3
Immunohistochemical Analysis of Collagen II
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Constructs were first, fixed with 4% paraformaldehyde overnight on the 14th day. Next, rinsed with PBS, dehydrated, and then embedded in paraffin. The samples were sectioned at 4 μm, deparaffinized, and rehydrated. To retrieve antigen, the sections were incubated in 0.1% pepsin for 15 min at 37 °C, and then blocking was carried out with 3% BSA. Polyclonal goat anti-human COL2A1 antibody (24128; Abcam) was consumed overnight at 4 °C, which was followed by incubation with 10 μg/mL fluorescein isothiocyanate (FITC) rabbit anti-goat secondary antibody (31509; Invitrogen, USA) for 1 h at room temperature. The DNA was labeled with 4′, 6-diamidino-2- phenylindole (DAPI). Negative controls without primary antibodies were prepared. Images were taken using an Axiovert 200M fluorescence microscope (Carl Zeiss Light Microscopy, Germany). The positive cells were counted to determine COL II level.
4
Immunofluorescent Staining of EGFR
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Cells were plated onto glass coverslips and allowed to grow to 70% confluence. Cells were washed with PBS and fixed in 4% PFA for 10 min at RT. The wells were further washed twice in PBS for 5 min each time and blocked in 10% (v/v) FBS for 1 h. Cells were then incubated with 1 µg/mL of primary anti-human EGFR goat polyclonal antibody (#AF231, R&D Systems) or a biotinylated anti-human IgG1 rabbit monoclonal antibody (#31-1019-02, 2BScientific) as a control in a humidified chamber for 1 h at RT. FITC rabbit anti-goat secondary antibody (#31509, Invitrogen) or streptavidin–FITC was added following 3 washes of 5 min each in PBS and counterstained using a Prolong Gold Antifade Mount nuclear stain (#P36930, ThermoFisher Scientific, Eugene, OR, USA). The stained cells were imaged using a Zeiss microscope (Jena, Germany) using filters for DAPI (acquisition time, 20 ms) and FITC (175 ms).
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