Anti foxp3 alexa fluor 488

Manufactured by Thermo Fisher Scientific

The Anti-Foxp3 Alexa Fluor 488 is a fluorescently labeled antibody that specifically binds to the Foxp3 transcription factor. Foxp3 is a key regulator of regulatory T cell (Treg) development and function. The Alexa Fluor 488 dye provides a green fluorescent signal that can be detected using flow cytometry or fluorescence microscopy.

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Alexa Fluor 488 (6806 citations) Alexa 488 (1863 citations) Anti-Foxp3 (119 citations) Alexa Fluors (21 citations) Alexa Fluor 488 anti (6 citations) FOXP3 Alexa 488 (4 citations) Anti-FOXP3-Alexa Fluor 488 (clone FJK-16s, (2 citations)

3 protocols using anti foxp3 alexa fluor 488

Multiparametric Flow Cytometry Panel

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The following antibodies were used for flow cytometric stainings anti-TACI PE (clone 1A1), anti-CD19 APC-Cy7, anti-CD27 PerCP-Cy5.5 or APC, anti-CD10 PE-Cy7, anti-IgM FITC, anti-CD21 APC, anti-CD69 PE-Cy7, anti-CD86 PE, anti-CD4 APC-Cy7, anti-CD25 PECy7, anti-CD127 PerCP-Cy5.5, anti-CD45RO Pacific Blue, anti-CXCR5 PerCP-Cy5.5, anti-PD-1 PE-Cy7, anti-CD25 PE, anti-CD25 PE-Cy7 (all from BioLegend, San Diego, Calif), anti-CD3 eFluor 605NC, anti-CD21 BD Horizon V450 (Becton Dickinson) and goat polyclonal anti-TACI biotin (R&D Systems). Intracellular staining with anti-Foxp3 Alexa Fluor 488 and anti-BCL6 PE (eBioscience, San Diego, Calif) was performed using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience), in accordance with the manufacturer's instructions.

Romberg N., Virdee M., Chamberlain N., Oe T., Schickel J.N., Perkins T., Cantaert T., Rachid R., Rosengren S., Palazzo R., Geha R., Cunningham-Rundles C, & Meffre E. (2015). TNFRSF13B hemizygosity reveals TACI haploinsufficiency at later stages of B-cell development. The Journal of allergy and clinical immunology, 136(5), 1315-1325.

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Flow Cytometric Immunophenotyping of Lymphocytes

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The following antibodies were used for flow cytometric stainings: anti-CD19 APC-Cy7, anti-CD27 PerCP-Cy5.5, anti-CD21 Pacific Blue, anti-CD4 APC-Cy7, anti-CD25 PE- Cy7, PE or PE Dazzle, anti-CD127 PerCP-Cy5.5 or APC, anti-CD45RO AF-700, anti-CXCR5 Pacific Blue, anti-PD-1 PE-Cy7 (all from BioLegend), anti-CD3 eFluor 605NC (eBioscience) and anti-IgG APC (Becton Dickinson). Intracellular staining with anti-Foxp3 Alexa Fluor 488 (eBioscience) or APC (Biolegend) was performed using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience) in accordance with manufacturer instructions.

Romberg N., Le Coz C., Glauzy S., Schickel J.N., Trofa M., Nolan B.E., Paessler M., Xu M.L., Lambert M., Lakhani S.A., Khokha M.K., Jyonouchi S., Heimall J., Takach P., Maglione P.J., Catanzaro J., Hsu F.I., Sullivan K.E., Cunningham-Rundles C, & Meffre E. (2018). CVID patients with autoimmune cytopenias exhibit hyperplastic yet inefficient germinal center responses. The Journal of allergy and clinical immunology, 143(1), 258-265.

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Phenotypic Analysis of Immune Cells

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Immune cells were treated with anti-CD16/32 for 10–15 min before being stained to block Fc receptors. Cells were then stained with anti-CD45 pacific blue, anti-CD4 APC-Fire/BV510, anti-CD8 PerCP-Cy5.5, anti-GITR-APC, anti-CD39-PE, anti-OX40-PE-Cy7, anti-CD11c BV510, anti-CD11b APC-Fire, anti-CD206 PerCP-Cy5.5, anti-GITRL-PE, or anti-Gr1 PE-Cy7 (all from BioLegend) at room temperature for 20–30 min. Staining for intracellular Foxp3 (Treg cell marker) involved fixing, permeabilizing, and staining cells with an anti-Foxp3 AlexaFluor 488 for 30 min at room temperature (eBioscience). Samples were collected and analyzed on an LSRII or a Gallios (BD Biosciences) flow cytometer and analyzed with FlowJo software.

Schoenhals J.E., Cushman T.R., Barsoumian H.B., Li A., Cadena A.P., Niknam S., Younes A.I., Caetano M.D., Cortez M.A, & Welsh J.W. (2018). Anti-glucocorticoid-induced Tumor Necrosis Factor–Related Protein (GITR) Therapy Overcomes Radiation-Induced Treg Immunosuppression and Drives Abscopal Effects. Frontiers in Immunology, 9, 2170.

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