Immunofluorescence staining was used to detect AAMs and Tregs on serial sections as described [25] (link). Tregs were detected using biotinylated anti-FoxP3 (FJK16s; eBioscience) with streptavidin-cyanin 3 (Jackson ImmunoResearch) and FITC-anti-CD4. Macrophages were stained with CD11c (BDPharmingen) for conventional M1 macrophages and polyclonal rabbit IgG anti-CD206 (Santa Cruz) followed by cyanin 3-labeled anti-rabbit antibody (Jackson) to identify AAMs (M2 macrophages). Images were acquired on an Olympus FluoView 500 BX51 confocal microscope and FluoView software version 4.3.
Biotinylated anti foxp3 fjk16s
Manufactured by Thermo Fisher Scientific
Biotinylated anti-FoxP3 (FJK16s) is a reagent used in flow cytometry and immunohistochemistry applications. It is a monoclonal antibody that specifically binds to the FoxP3 transcription factor, which is a marker for regulatory T cells. The antibody is conjugated with biotin, which allows it to be detected using streptavidin-based detection systems.
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2 protocols using biotinylated anti foxp3 fjk16s
1
Immunofluorescence Staining for AAMs and Tregs
2
Multiparameter Immunofluorescence Imaging of Lymphoid Tissues
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5-µm-thick frozen sections of OCT-embedded spleens, peripheral lymph nodes, small intestines, or Peyer’s patches were fixed with 3.2% formaldehyde (Polysciences, Inc.), permeabilized in 90% methanol, and stained with anti-CD5 (53–7.3; BioLegend) conjugated to Alexa Fluor 488, biotinylated anti-Foxp3 (FJK-16s; eBioscience) visualized with Alexa Fluor 647–conjugated streptavidin, anti-pStat5 (pY694; D47E7; Cell Signaling Technology) visualized with Alexa Fluor 555–conjugated anti–rabbit IgG F(ab′)2 (Cell Signaling Technology), or biotinylated anti–MAdCAM-1 (MECA-367; eBioscience) visualized with Alexa Fluor 555–conjugated streptavidin. Images were acquired on a confocal microscope (TCS-SP5 II; Leica) and analyzed using ImageJ software (National Institutes of Health).
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