Pe anti foxp3 antibody

Manufactured by BioLegend

Sourced in United States

The PE anti-FOXP3 antibody is a fluorescence-conjugated antibody specific for the transcription factor FOXP3. FOXP3 is a key regulator of T-regulatory cell development and function. The PE-conjugated anti-FOXP3 antibody can be used for the identification and analysis of FOXP3-expressing cells by flow cytometry.

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2 protocols using pe anti foxp3 antibody

Multiparametric Flow Cytometry Analysis

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Up to 1.10⁶ cells were labelled in staining solution (PBS, Foetal Calf Serum 2%, 2mM EDTA). Antibodies used were: APC/Cy7 anti-CD3e, PE/Cy7 anti-CD4, APC anti-CD25, PE anti-CD44, FITC anti-CD62L (all from Biolegend). Dead cells were stained either with 7-AAD (Biolegend) or Livedead cell viability assay (Life Technologies). Cells were fixed using fixation solution 20 minutes at room temperature (Biolegend). For regulatory T cells labelling, FOXP3 Perm/Fix solution was use with PE anti-FOXP3 antibody (Biolegend) according to manufacturer’s instructions. Intracellular IFNγ staining was performed using permeabilisation buffer following manufacturer’s instructions and FITC anti-IFNγ antibody and APC anti-IL4 antibody (Biolegend). Data were acquired on a BD FACSCanto II flow cytometer (BD) and analyzed employing FlowJo software (Tree Star).

Bonaccorsi-Riani E., Danger R., Lozano J.J., Martinez-Picola M., Kodela E., Mas-Malavila R., Bruguera M., Collins H.L., Hider R.C., Martinez-Llordella M, & Sanchez-Fueyo A. (2015). Iron Deficiency Impairs Intra-Hepatic Lymphocyte Mediated Immune Response. PLoS ONE, 10(8), e0136106.

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Multicolor Flow Cytometry Profiling of T Cell Subsets

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A single-cell suspension was stimulated for 4 hours at 37°C with phorbol myristic acetate (50 ng/mL), ionomycin (1 mg/mL), and brefeldin A (1 mg/mL) (Sigma-Aldrich, St. Louis, MO, USA); then, cells were fixed, permeabilized overnight, and intracellularly stained with antibodies. Tregs were detected directly without stimulations. Cell surface proteins were stained with PE-anti-CD4 (cat. 116006; BioLegend, San Diego, CA, USA) or FITC-anti-CD4 (cat. 130308; BioLegend) antibodies for 20 minutes at 4°C, and intracellular cytokines were stained with APC-anti-IFN-γ antibody (cat. 505810; BioLegend) for Th1 cells, FITC-anti-IL-17 antibody (cat. 506908; BioLegend) for Th17 cells, or PE-anti-Foxp3 antibody (cat. 12-5773-82; eBioscience, San Diego, CA, USA) for Tregs for 1 hour at room temperature. Stained cells were collected on a flow cytometer (FACS Calibur; BD Biosciences, San Jose, CA, USA). Data were analyzed with FlowJo software (TreeStar, Ashland, OR, USA).

Guo X., Dang W., Li N., Wang Y., Sun D., Nian H, & Wei R. (2022). PPAR-α Agonist Fenofibrate Ameliorates Sjögren Syndrome–Like Dacryoadenitis by Modulating Th1/Th17 and Treg Cell Responses in NOD Mice. Investigative Ophthalmology & Visual Science, 63(6), 12.

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