Anti mouse foxp3

Manufactured by BioLegend

Sourced in United States

Anti-mouse FoxP3 is a lab equipment product designed to detect and quantify the expression of the FoxP3 transcription factor in mouse samples. FoxP3 is a key marker for regulatory T cells (Tregs) and is essential for their development and function. This product can be used in various applications, such as flow cytometry and immunohistochemistry, to study the role of Tregs in different biological and disease processes.

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4 protocols using anti mouse foxp3

Comprehensive Immune Cell Profiling

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The following antibodies were used for the present study.
Anti-mouse CD3, clone number 17A2, BioLegend, 05112-40-100, V450;
Anti-mouse CD4, clone number GK1.5, BioLegend, 100434, PerCP/Cyanine5.5;
Anti-mouse CD8, clone number 53-6.7, BioLegend, 100714, APC/Cyanine7;
Anti-mouse CD25, clone number 3C7, BioLegend, 101908, FITC;
Anti-mouse CD69, clone number H1.2F3, BioLegend, 104508, PE;
Anti-mouse CTLA-4, clone number UC10-489, BioLegend, 106305, PE;
Anti-mouse PD-1, clone number 29F.1A12, BioLegend, 135210, APC;
Anti-mouse CD19, clone number 1D3, eBioscience, 12-0193-82, PE;
Anti-mouse CD45, clone number 30F11, BioLegend, 103122, Alexa Fluor 488;
Anti-human CD45, clone number HI30, BioLegend, 304025, PerCP;
Anti-mouse IFNγ, clone number XMG1.2, eBioscience, PE;
Anti-mouse IL-17A, clone number TC11-18H10.1, BioLegend, Alexa Fluor 647;
Anti-mouse IL-13, clone number 13A, BioLegend, PE/Cyanine7;
Anti-mouse Foxp3, clone number MF14, BioLegend, PE;
Anti-mouse TNFα, clone number MP6-XT22, BioLegend, Alexa Fluor 488;
Purified Anti-mouse IFNγ antibody, clone number XMG1.2;
Purified anti-mouse IL-4 antibody, clone number 11B11.

Yang F.M., Shen L., Fan D.D., Chen K.H, & Lee J. (2022). DMGV Is a Rheostat of T Cell Survival and a Potential Therapeutic for Inflammatory Diseases and Cancers. Frontiers in Immunology, 13, 918241.

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Tumor-Infiltrating Lymphocyte Analysis Protocol

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Tumor-infiltrating lymphocytes were analyzed as previously described^{36 (link),37 (link)}. Briefly, 4 days after the last injection, the tumor masses were harvested from euthanized mice and weighed. Then, the tumor tissues were minced and passed through a 70-μm pore filter. The separated single cells were stained with the following antibodies: anti-mouse CD3, anti-mouse CD4, anti-mouse CD8, anti-mouse Foxp3, anti-mouse IFN-γ and anti-mouse TNF-α (BioLegend). Intracellular staining of tumor-infiltrating lymphocytes was conducted according to the manufacturer’s protocol. The staining results were analyzed using FlowJo software (TreeStar Inc.).

Hong J., Jung M., Kim C., Kang M., Go S., Sohn H., Moon S., Kwon S., Song S.Y, & Kim B.S. (2023). Senescent cancer cell-derived nanovesicle as a personalized therapeutic cancer vaccine. Experimental & Molecular Medicine, 55(3), 541-554.

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Mouse Spleen Cell Immunophenotyping

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The antibodies used for flow cytometric analysis include FITC anti-mouse CD3, Pacific Blue anti-mouse CD4, PerCP anti-mouse CD8a, PE/Cyanine 7 anti-mouse CD19, APC anti-mouse CD25, PE anti-mouse FoxP3 and their corresponding isotype controls. All of them were obtained from Biolegend (San Diego, CA, USA).
The mouse spleen cells were suspended in RPMI 1640 (HyClone, Logan, UT, USA) medium with 10% fetal calf serum (Hangzhou Sijiqing Biological Engineering Materials) at a final concentration of 1×10⁷cells/mL. Splenocytes were then suspended in a flow cytometry staining buffer (Biolegend), stained with surface antibodies (anti-mouse CD3, anti-mouse CD4, anti-mouse CD8, anti-mouse CD19 and anti-mouse CD25) for 30 min, incubated with permeabilization buffer (Biolegend) at room temperature for 1 hour and stained with the intracellular antibody (anti-mouse FoxP3) according to the manufacturer’s instructions. Flow cytometric analysis was performed using a flow cytometry machine (Beckmancount, Shanghai, China).

Ye Z., Wu H., Chen X., Xie R., Zhang D., Sun H., Wang F., Li Z., Xia Q., Chen L, & Chen T. (2024). Puerarin inhibits inflammation and oxidative stress in female BALB/c mouse models of Graves’ disease. Translational Pediatrics, 13(1), 38-51.

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FoxP3 Acetylation Quantification in Tregs

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Viable Tregs and Tcon cells isolated as described above were subjected to proximity ligation assay as described [62 (link)] to measure the acetylation of FoxP3. Briefly, sorted cells (1–5×10⁴ cells) were centrifuged onto glass slides (Cytospin), fixed with periodate-lysine-paraformaldehyde, permeabilized with 0.2% Triton X-100, and stained with anti-mouse FoxP3 (Clone:MF14, BioLegend) and anti-rabbit acetylated lysine (Clone:RM101, Abcam). Slides were subsequently treated with DuoLink in situ PLA probes kit (Sigma) as per the manufacturer’s instructions. Nuclei were counterstained with DAPI. Cells were imaged using a ZOE fluorescent microscope (BioRad), and an Axioscope 5 (Zeiss) and the proportions of DAPI+ cells positive for the FoxP3-specific acetylated lysine signal was found via ImageJ.

Rosario S.R., Smith RJ J.r., Patnaik S.K., Liu S., Barbi J, & Yendamuri S. (2022). Altered acetyl-CoA metabolism presents a new potential immunotherapy target in the obese lung microenvironment. Cancer & Metabolism, 10, 17.

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