Anti mouse cd25

Manufactured by BioLegend

Sourced in United States

Anti-mouse CD25 is a monoclonal antibody that binds to the CD25 antigen expressed on the surface of activated T cells, regulatory T cells, and other immune cell types in mice. It can be used for flow cytometry and other immunological applications to identify and characterize CD25-positive cell populations.

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Lab products found in correlation

Anti mouse cd4, by BioLegend (4 mentions) Anti mouse cd3, by BioLegend (3 mentions) Anti mouse cd8, by BioLegend (3 mentions) Anti mouse cd69, by BioLegend (2 mentions) Anti mouse foxp3, by BioLegend (2 mentions) Anti mouse tnf α, by BioLegend (2 mentions) Anti mouse cd62l, by BioLegend (2 mentions) Fcs express software, by De Novo Software (2 mentions) Foxp3 staining kit, by Thermo Fisher Scientific (2 mentions) Bv605, by BioLegend (2 mentions) Pe anti mouse lap, by BioLegend (2 mentions) Cd4 efluor450, by Thermo Fisher Scientific (2 mentions) Lsrii flow cytometer, by BD (2 mentions) Anti mouse pd 1, by BioLegend (1 mentions) Anti mouse cd19, by Thermo Fisher Scientific (1 mentions) Anti mouse cd45, by BioLegend (1 mentions) Anti human cd45, by BioLegend (1 mentions) Anti mouse ifn γ, by Thermo Fisher Scientific (1 mentions) Anti mouse il 17a, by BioLegend (1 mentions) Anti mouse ifn γ antibody, by BioLegend (1 mentions)

9 protocols using anti mouse cd25

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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Mouse Model of Acute GVHD

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Mouse models of aGVHD are well documented [24 (link), 25 (link)]. Recipient BALB/c mice were irradiated with 780 cGy of irradiation using a Cesium 137 irradiator in 2 sessions 4 h apart. Donor CD45.1⁺ C57BL/6 mice were sacrificed on the day of transplant and bone marrow (BM) and spleen dissected out. Femur and tibia was used for marrow isolation. Spleen was sorted for T cells by selecting for cells that were negative for Peridinin-Chlorophyll-Protein/Cyanine5.5 (PerCP/Cy5.5) conjugated anti–mouse B220 (Biolegend, Cat. #103236) and anti–mouse CD25 (Biolegend, Cat. #102030) using MoFlo XDP. Recipients were administered 1 × 10⁶ of BM and 1 × 10⁶ sorted T cells through retro-orbital injections. Cohorts receiving ILC2s received 1 × 10⁶ of ILC2s. All animal experiments were performed according to protocols approved by the Institutional Animal Care and Use Committee policies at the Oklahoma Medical Research Foundation and in compliance with the ARRIVE guidelines.

Srinivasan A., Bajana S., Pankow A., Yuen C., Shah R.K, & Sun X.H. (2021). Type 2 innate lymphoid cells from Id1 transgenic mice alleviate skin manifestations of graft-versus-host disease. BMC Immunology, 22, 46.

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Flow Cytometric Analysis of Immune Cell Profiles

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Flow cytometry was used for analyzing the toxic effects of HCQ on lymphocytes with Zombie NIR™ Dye (Biolegend, San Diego, CA), and the frequencies of naïve CD4+T (CD4+CD62L+CD44-), Th1 (CD4+IFN-γ+), Th17 (CD4+IL-17A+), T regulator (Tregs) (CD4+CD25+Foxp3+) cells, and the expression of LOX-1 in RVECs. The cells were incubated with Fc block (clone 2.4G2, Bio Xcell) and stained with following antibodies from BioLegend: anti-mouse CD3 (BV421), anti-human CD3 (BV421), anti-human CD8 (PE), anti-human LOX-1 (APC), anti-mouse CD4 (Percp-cy5.5), anti-mouse CD25 (PE-cy7), anti-mouse CD44 (APC), anti-mouse CD62L (FITC), and anti-mouse CD69 (PE). For intracellular cytokine staining, the cells were stimulated with phorbol 12-myristate 13-acetate (PMA) (50 ng/mL; Sigma), ionomycin (500 ng/mL; Sigma), and Brefeldin A (1 µg/mL; Sigma) for 4–5 h. The The intracellular cytokines or transcription factor were stained with anti-human/mouse IFN-γ (BV786), anti-human/mouse IL-17A (BV650), and anti-human/mouse Foxp3 (FITC) after fixation and permeabilization. Data were analyzed using the FlowJo software 10.0 (Tree Star, Ashland, OR, USA).

Hu Y., Li Z., Chen G., Li Z., Huang J., Huang H., Xie Y., Chen Q., Zhu W., Wang M., Chen J., Su W., Chen X, & Liang D. (2022). Hydroxychloroquine Alleviates EAU by Inhibiting Uveitogenic T Cells and Ameliorating Retinal Vascular Endothelial Cells Dysfunction. Frontiers in Immunology, 13, 859260.

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Activation of OT-I CD8+ T Cells by Infected DCs

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HI rWR (A3 or F17 where indicated) or UV-attenuated WR-mini-OVA was used to infect either MutuDCs, 293A cells, or 293KbC2 cells (57 (link)) for 2 h with rocking at an MOI of 10 before the inoculum was replaced with IMDM-10. Splenocytes from a naive OT-I mouse were harvested and subjected to CD8α-negative enrichment (Miltenyi Biotech, 130-096-543). The eEnrichment efficiency was checked by flow cytometry, and samples were at least 85% CD8α⁺ Vα2⁺ T cells. OT-I CD8⁺ T cells were added to V-bottom plates in D10 with β-mercaptoethanol, and infected cells were added in the same medium at a ratio of 1:2 (infected cell target to OT-I). After 24 h, the cells were labeled with anti-mouse CD11c-FITC (BioLegend, clone N418), anti-mouse CD8α-PE-Cy7 (BioLegend, clone 53.67), anti-mouse TCR Vα2-APC (BioLegend, clone B20.1), anti-mouse CD69-PerCP-Cy5.5 (BioLegend, clone H1.2F3), and anti-mouse CD25 (BioLegend, clone 3C7) antibodies diluted in PBS with 2% FBS. Events were gated sequentially on SSC-A × FSC-A (lymphocytes), FSC-H × FSC-W (single cells), SSC-H x SSC-W (single cells), SSC-A × FITC/GFP^– (MutuDC exclusion), PE-Cy7 × APC (CD8⁺ Vα2⁺ cells), and PE × PerCP-Cy5.5 (CD25⁺ CD69⁺ activated cells) to determine the percentages of CD8⁺ Vα2⁺ T cells that upregulate activation markers.

Croft S., Wong Y.C., Smith S.A., Flesch I.E, & Tscharke D.C. (2020). Surprisingly Effective Priming of CD8+ T Cells by Heat-Inactivated Vaccinia Virus Virions. Journal of Virology, 94(20), e01486-20.

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Immunophenotyping and FoxP3 Detection in Murine Tumor Models

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PE mesh (fiber diameter: 86 µm, pitch: 125 µm/163 µm) was purchased from Semitec Corp. (Osaka, Japan). Antibodies (anti-mouse CD25, anti-human CD25, anti-mouse CD4, fluorescein isothiocyanate [FITC]-labeled anti-mouse CD25, and allophycocyanin [APC]-labeled anti-mouse CD4) were purchased from BioLegend, Inc. (San Diego, California, USA). Monoclonal anti-FoxP3 antibodies were purchased from Thermo Fisher Scientific (Waltham, Massachusetts, USA). Immunostaining kit (POD conjugate anti-rat, for mouse tissue) was purchased from Takara Bio Inc. (Shiga, Japan). Simple Stain MAX-PO (Rat) and diaminobenzidine (DAB) Substrate Kit were purchased from Nichiren Bioscience Inc. (Tokyo, Japan). B16 melanoma cells were purchased from the JCRB cell bank (Osaka, Japan). C57BL/6 mice were purchased from Sankyo Lab Service Corp. (Tokyo, Japan).

Kimura T., Tokunaga R., Hashimoto Y., Nakamura N, & Kishida A. (2021). Tumor growth suppression by implantation of an anti-CD25 antibody-immobilized material near the tumor via regulatory T cell capture. Science and Technology of Advanced Materials, 22(1), 607-615.

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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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Murine Immune Cell Profiling

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Murine blood samples were pre-incubated with rat-anti mouse CD16/32 (Pharmigen, Franklin Lakes, NJ) and then stained with a mixture of efluor 450 CD4 (eBioscience, Waltham, MA), BV605, anti-mouse CD25 (Biolegend, San Diego, CA) and PE-anti mouse LAP (Biolegend). Intracellular staining for forkhead box P3 (FoxP3) was performed with FoxP3 staining kit (eBioscience). Data were collected on LSR II flow cytometer (BD Bioscience, San Jose, CA) and analyzed with FCS express software (De Novo Software, Glendale, CA).

Kwon K., Sherman A., Chang W., Kamesh A., Biswas M., Herzog R.W, & Daniell H. (2017). Expression and assembly of largest foreign protein in chloroplasts: oral delivery of human FVIII made in lettuce chloroplasts robustly suppresses inhibitor formation in haemophilia A mice. Plant Biotechnology Journal, 16(6), 1148-1160.

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Murine Regulatory T Cell Immunophenotyping

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Murine blood samples were pre‐incubated with rat anti‐mouse CD16/32 (Pharmingen, Franklin Lakes, NJ) and then stained with a mixture of eFluor 450 CD4 (eBioscience, Waltham, MA), BV605, anti‐mouse CD25 (Biolegend, San Diego, CA) and PE anti‐mouse LAP (Biolegend). Intracellular staining for forkhead box P3 (FoxP3) was performed with FoxP3 staining kit (eBioscience). Data were collected on LSR II flow cytometer (BD Bioscience, San Jose, CA) and analysed with FCS express software (De Novo Software, Glendale, CA).

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Spleen Immunophenotyping by Flow Cytometry

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The spleen was isolated from experimental mice, then followed by homogenisation for intracellular and extracellular immunostaining procedure as similar as our previous study (Putra et al. 2015 (link)). Several antibodies were used such as anti-mouse CD4, anti-mouse CD8, anti-mouse CD62L, anti-mouse CD25, anti-mouse TNF-α, and anti-mouse IFN-γ (Biolegend). Flow cytometry analysis was accomplished by BD FACS CaliburTM (BD Bioscience).

Putra W.E, & Rifa’i M. (2020). Assessing the Immunomodulatory Activity of Ethanol Extract of Sambucus javanica Berries and Leaves in Chloramphenicol-Induced Aplastic Anemia Mouse Model. Tropical Life Sciences Research, 31(2), 175-185.

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