Cd25 bv510

Manufactured by BD

Sourced in China, United States

The CD25-BV510 is a fluorescently-labeled antibody that binds to the CD25 antigen, which is expressed on the surface of activated T cells and regulatory T cells. This product is intended for use in flow cytometry applications to identify and quantify these cell populations.

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3 protocols using cd25 bv510

Immunoprofiling of Tumor Microenvironment

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Antibodies for immunohistochemical staining: anti-CD8α (D4W2Z, 98941), Foxp3 (D6O8R, 12653S), F4/80 (D2S9R, 70076S), CD206 (E6T5J, 24595S) were obtained from Cell Signaling Technology (Danvers, MA). TUNEL Reagent Test Kit (11684795910) was purchased from Roche (Shanghai, China).
Antibodies for Flow Analysis: CD8α-PerCP-Cy5.5 (53–6.7, 561109), CD25-BV510 (PC61, 563037), CD279 (PD-1)-BV605 (RMP1–30, 748267), CD3-BV650 (145-2C11, 564378), IL-10-BV711 (JES5-16E3, 564081), IL-17A-BV786 (TC11-18H10, 564171), CD45-BUV395 (30-F11, 565967), CD366 (Tim-3)-PE (5D12, 566346), STAT3-PE-CF594 (4/P-STAT3, 562673), CD4-PE-Cy7 (GK1.5, 563933), CD206-Alexa Fluor® 647 (MR5D3,565250), F4/80-BV510 (T45-2342,743280) were all obtained from BD Bioscience (San Jose, CA, USA). Foxp3-Alexa Fluor® 647 (150D, 320014), TGF-β1-FITC (TW7-16B4, 141414), Granzyme B-BV421(QA18A28, 396414) were purchased from BioLegend (San Diego, CA, USA). Fixable viability Dye eFluor^TM 780 (65-0865-14) were purchased from eBioscience^TM (San Diego, CA, USA).

Huang L., Zhao Y., Shan M., Wang S., Chen J., Liu Z, & Xu Q. (2023). Targeting crosstalk of STAT3 between tumor-associated M2 macrophages and Tregs in colorectal cancer. Cancer Biology & Therapy, 24(1), 2226418.

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Multiparameter Flow Cytometry Immunophenotyping

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For surface staining, peripheral blood mononuclear cells (PBMCs) were incubated with directly conjugated mAbs for 30 min at 4 °C and then washed before analysis. Antibodies used were anti-human CD3-BV786 or CD3-APC, CD4-BV711, CD8-APC-H7, CD45RA-AF700, CD95-FITC, CD25 BV510, CD73 PE-CY7, CD11b AF700, CD45 BV786, CD14 BV711, CD56-PE-CF594, CD19-FITC, HLA-DR-PE, CD160-AF488 (BD Biosciences, San Diego, CA, USA), CCR7-BV421, PD-1-PE, 2B4-APC (BioLegend, San Diego, CA, USA), TIGIT-APC (Ebioscience, San Diego, CA, USA), LAG-3-AF700 (R&D Systems, Minneapolis, MN, USA) antibodies, and corresponding isotype controls. Data acquisition was performed on a LSR Fortessa flow cytometer (BD Biosciences), and data analysis was performed using FlowJo Software (Tree Star, Ashland, OR, USA).

Kong Y., Jia B., Zhao C., Claxton D.F., Sharma A., Annageldiyev C., Fotos J.S., Zeng H., Paulson R.F., Prabhu K.S, & Zheng H. (2019). Downregulation of CD73 associates with T cell exhaustion in AML patients. Journal of Hematology & Oncology, 12, 40.

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Comprehensive T Cell Subset Analysis

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For T cell subset analyses, 100 μL of whole blood was mixed with an optimal dilution of the following monoclonal antibodies: CD3-APC-CY7, CD4-PE, CD8-PerCP-Cy5.5, CD45RA-FITC, CCR7-PE-CF594, CD5-PE-CY7, CD27-APC, CD127-V450, CD25-BV510 (BD Bioscience). After adding all antibodies, the mixture was gently vortexed and incubated for 15 min in the dark at room temperature. Subsequently, 2 mL of 1× FACS lysing solution (BD Bioscience) was added and incubated for 10 min in the dark at room temperature. Finally, cells were washed 3 times with 200 μL washing buffer. Data was collected by a 9-colour Aria I flow cytometer (BD Bioscience) and analyzed with FACSDiva software (version 6.1.3, BD Bioscience). The following T cell subsets were determined (Mahnke et al., 2012; Schatorje et al., 2012)
Tem), and CD8 + transitional memory T cells (CD3 + CD8 + CD4 -CD45RA -CCR7 -CD27 + , CD8 + Ttm). The full-minus-one (FMO) principle was adopted for gating the T cell subsets. Absolute counts of each subgroup were calculated by multiplying the total lymphocyte counts with the percentages of each subset that were determined by flow cytometry.

Cao J., Xu X., Zhang Y., Zeng Z., Hylkema M.N, & Huo X. (2018). Increased memory T cell populations in Pb-exposed children from an e-waste-recycling area. The Science of the total environment, 616-617.

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