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Fitc conjugated anti il 17 antibodies

Manufactured by Thermo Fisher Scientific
Sourced in United States

FITC-conjugated anti-IL-17 antibodies are laboratory reagents used for the detection and analysis of interleukin-17 (IL-17) in biological samples. These antibodies are conjugated with fluorescein isothiocyanate (FITC), a fluorescent dye, to facilitate visualization and quantification of IL-17 in flow cytometry, immunohistochemistry, and other immunoassay applications.

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3 protocols using fitc conjugated anti il 17 antibodies

1

Intracellular Detection of IL-17 in Th17 Cells

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For intracellular detection of IL-17, CD4+ T cells were incubated with 25 ng/mL phorbol 12-myristate 13-acetate (PMA), 250 ng/mL ionomycin (Sigma-Aldrich), and monensin-containing GolgiStop (BD Biosciences, San Jose, CA, USA) for 4 h. The harvested cells were stained with PerCP-conjugated anti-CD4 antibodies (Biolegend, San Diego, CA, USA). After fixation with fixation/permeabilization solution, the cells were stained with 0.125 μg FITC-conjugated anti-IL-17 antibodies (eBioscience, San Diego, CA, USA) to determine the population of Th17 cells. All analyses were performed using a BD LSRII fortessa (BD Biosciences) and FACS DIVA version 10.0 (BD Biosciences).
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2

Immunostaining of T Cell Subsets

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For immunostaining, 7-μm tissue sections of spleens were used. To analyze the populations of T helper 17 cells, we used PE-conjugated anti-CD4 and fluorescein isothiocyanate (FITC)-conjugated anti-IL-17 antibodies (eBioscience; San Diego, CA, USA). To analyze the populations of regulatory T cells, the samples were stained with FITC-conjugated anti-CD4, APC-conjugated anti-CD25, and PE-conjugated anti-Foxp3 antibodies. The stained sections were observed using a Zeiss microscope (LSM 510 Meta; Carl Zeiss, Oberkochen, Germany) at × 400 magnification. Positive cells were counted, and the values were expressed as the mean ± standard deviation.
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3

Characterizing T and B Cell Subsets

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The cells used for analysis of the CD4+ IFNγ + (Th1), CD4+ IL-4+ (Th2), CD4+ IL-17+ (Th17), and IL-17-secreting CD19+ B cell (B17) populations were stimulated with PMA and ionomycin for 4 h using GolgiStop (BD Biosciences, Franklin Lake, NJ, USA). To quantify the Th1, Th2, Th17, and B17 cell populations, mouse splenocytes were immunostained using a PerCp5.5-conjugated anti-CD4 antibody (eBioscience, San Diego, CA, USA) for T cells and a PerCP5.5-conjugated anti-CD19 antibody for B cells. The cells were fixed and permeabilized using the Cytofix/Cytoperm Plus kit (BD Biosciences) following the manufacturer’s instructions, and then stained with APC-conjugated anti-IFNγ, PE-conjugated anti-IL-4, APC-conjugated anti-IL-10, and FITC-conjugated anti-IL-17 antibodies (eBioscience), respectively. All samples were analyzed using the Attune NxT Flow cytometer (Thermo Fisher Scientific, Waltham, MA, USA). The flow cytometry data were analyzed using FlowJo™ software (FlowJo, Ashland, OR, USA).
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