Cd25 apc pc61

Manufactured by BD

CD25-APC (PC61) is a fluorescent-labeled antibody that binds to the CD25 antigen, also known as the alpha chain of the interleukin-2 receptor (IL-2Rα). It is a commonly used marker for the identification and analysis of regulatory T cells (Tregs) in flow cytometry applications.

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CD25-APC (96 citations) CD25 (PC61; (18 citations) Anti-CD25 APC (PC61; (3 citations) Anti-mouse CD25 APC (PC61, (2 citations) APC Rat Anti-Mouse CD25 (Clone PC61, (2 citations) APC-conjugated anti-CD25 (clone PC61); (2 citations) APC)-conjugated CD25 (PC61; (2 citations)

2 protocols using cd25 apc pc61

Comprehensive Flow Cytometry Analysis

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Flow cytometry was performed with CantoII (BD) with monoclonal antibodies CD4-FITC (RM4-5, eBioscience), CD4-PerCP (RM4-5, BD Bioscience), CD25-APC (PC61, BD Bioscience), CD62L-FITC (MEL-14, BD Bioscience), CD69-FITC (H1.2F3, BD Bioscience) and FoxP3-PE (FJK-16, eBioscience). For Fig 1A, whole blood cells were obtained via the submandibular vein 7 days after administration of anti-CD25 antibody (which would be the time point for peptide immunization). Red blood cells were lysed with ACK (Ammonium-Chloride-Potassium) buffer. Remaining cells were stained for CD25 and the percentage of CD25⁺ cells was determined for the entire population of blood cells. For Fig 2, single cell suspensions of splenocytes were stained for CD4 in combination with CD25, FoxP3, CD69, or CD62L.

van Herwijnen M.J., van der Zee R., van Eden W, & Broere F. (2015). In Vivo Induction of Functionally Suppressive Induced Regulatory T Cells from CD4+CD25- T Cells Using an Hsp70 Peptide. PLoS ONE, 10(6), e0128373.

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Multiparametric Flow Cytometry Analysis

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The following antibodies from BD Biosciences (Franklin Lakes, NJ) were used for staining: CD8-FITC (53.67), CD8-APC (KT15), Foxp3-PE (FJK-16s), CD3-AF700 (17A2), CD44-PerCP-Cy5.5 (IM7), CD4-FITC (RM4– 5), CD62L-BV421 (MEL-14), CD25-APC (PC61), Ki67-PE-Cy7 (SolA15), CTLA-4-PE (UC10–4F10–11), PD-1-BV421 (RMP1–30), granzyme B-PE (NGZB), OX40-BV711 (OX86), GITR-BV510 (DTA-1). MHC class I-restricted (H2-Db) PE-labeled CEA-tetramer (sequence: EAQNTTYL) was purchased from MBL International Corporation (Woburn, MA). Live/Dead fixable aqua stain and transcription factor staining buffer set were purchased from Thermo Fisher (Waltham, MA). Flow cytometry was performed on BD LSRFortessa or BD FACSVerse (BD Biosciences) and analyzed using FlowJo v.9.7.6 or v.10.5.3 (TreeStar). Cell viability was examined using trypan blue staining prior to data acquisition. Live cells were gated via forward and side scatter. Isotype control staining was <5% for all samples analyzed.

Fabian K.P., Malamas A.S., Padget M.R., Solocinski K., Wolfson B., Fujii R., Sater H.A., Schlom J, & Hodge J.W. (2020). Therapy of Established Tumors with Rationally Designed Multiple Agents Targeting Diverse Immune-Tumor Interactions: Engage, Expand, Enable. Cancer immunology research, 9(2), 239-252.

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