Additional male and female DOCA rats (n=5–8) were randomized to receive weekly intraperitoneal (ip) injections of anti-CD25 antibody for 3 weeks (1 mg/ml; eBioscience cat # 16-0390-85) or anti-IgG (control). The anti-CD25 antibody targets the CD25 surface marker on Tregs to decrease circulating Tregs20 (link)–22 (link). Pilot studies determined the dose of anti-CD25 to decrease Tregs in male and female SD rats (Online Supplement ). Based on pilot studies, rats were injected with 250 μg of anti-CD25 every seven days for 3 weeks to decrease Tregs.
Anti cd25 antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Anti-CD25 antibody is a laboratory reagent used for the detection and analysis of CD25-expressing cells. CD25 is a cell surface marker expressed on activated T cells, regulatory T cells, and other immune cell types. The antibody can be used in various immunological techniques, such as flow cytometry and immunohistochemistry, to identify and study these cell populations.
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Lab products found in correlation
3 protocols using anti cd25 antibody
1
Treg Depletion in DOCA-Induced Hypertension
2
Immunophenotyping of Treg and Th17 Cells
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For the analysis of the Treg cells, the cells were stained with surface marker α chain of the high-affinity IL-2 receptor (CD25) in PBS containing 1% BSA and anti-CD25 antibody (eBioscience, USA). Foxp3 staining was then performed in accordance with the manufacturer’s instructions (eBioscience, USA). For the analysis of the Th17 cells, the cells were stimulated with leukocyte activation cocktail (BD Pharmagen, USA) for 6 hours, and the cells were then collected and re-suspended in a fixation/permeabilization solution (BD Pharmagen, USA), and stained with anti-IL-17A antibody (BD Pharmagen, USA).
3
Activation and Characterization of CD8+ T Cells
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To activate CD8þ T cells, the isolated CD8þ T cells were resuspended in RPMI 1640 (GIBCO, Grand Island, New York) supplemented with 10% fetal bovine serum, 50 IU∕ml penicillin, and 50 lg ∕ml streptomycin (Hyclone, Logan, Utah) to the desired cell density (2 × 10 5 cells/48 well) on plates coated with anti-CD3 antibody (eBioscience, San Diego, California) and soluble anti-CD28 antibody (eBioscience, San Diego California). After three days, CD8þ T cells were treated with anti-CD25 antibody (eBioscience, San Diego, California), resuspended in flow cytometry staining buffer, and immediately subjected to flow cytometric analysis. To confirm the presence of activated mature CD8þ T cells, some three-day old CD8þ T cells were treated with anti-CD25 antibody/anti-Fas antibody (eBioscience, San Diego California) and anti-CD25 antibody/ anti-Annexin-V antibody (Sigma Aldrich, St. Louis, Missouri) for 30 min. Flow cytometric data were acquired using a FACScalibur flow cytometer.
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