Cd25 apc

Manufactured by Beckman Coulter

Sourced in United States

The CD25-APC is a flow cytometry reagent that detects the expression of the CD25 antigen on the surface of cells. CD25 is the alpha chain of the interleukin-2 receptor, which is a marker for T-cell activation. The CD25-APC reagent is conjugated with the fluorescent dye Allophycocyanin (APC), allowing for the identification and quantification of CD25-positive cells in a sample by flow cytometry.

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2 protocols using cd25 apc

Multicolor Flow Cytometry for T and B Cell Subsets

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PBMC were isolated from peripheral blood by Ficoll gradient and frozen for batched analysis. We designed six multicolor flow cytometry panels to quantify 60 T cell subsets along with two B cell subsets, and calculated the CD4⁺/CD8⁺ T cell ratio (Supplementary Table 1). The following fluorochrome-conjugated anti-human antibodies were used from BD Biosciences (San Jose, CA): CD3-FITC, CD3-PerCP-Cy5.5, CD4-APC, CD4-APC-Cy7, CD8-BV510, CD45RO-FITC, CD45RA-APC, CD45RA-APC-Cy7, CCR4-PE, CD27-PE, CD28-BV421, CD138-BV421, CCR6-BV421, CXCR3-PE, CCR7-A700, IL-17-BV786, IFN-γ-PE-Cy7, iso IgG1k-FITC, iso IgG1k-PE-Cy7, iso IgG2bk-APC, iso IgG1k-APC-Cy7, and iso IgG1k-BV510; from Biolegend (San Diego, CA): CD127-FITC, CD27-APC, CD57-PerCp-Cy5.5, CD19-BV510, PD-1-APC-Cy7, CXCR5-FITC, and TNFα-FITC; from eBioscience (San Diego, CA): CD4-PE-Cy7, and IL-2-PE; from Miltenyi Biotec (San Diego, CA): CD25-APC and KLRG1-PE; from Beckman Coulter (Brea, CA): CD38-PE-Cy7.

Hartzell S., Bin S., Cantarelli C., Haverly M., Manrique J., Angeletti A., Manna G.L., Murphy B., Zhang W., Levitsky J., Gallon L., Yu S.M, & Cravedi P. (2020). Kidney Failure Associates With T Cell Exhaustion and Imbalanced Follicular Helper T Cells. Frontiers in Immunology, 11, 583702.

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Quantification of Th17 and Treg Cells

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Absolute count of Th17 and Treg cells was performed by flow cytometry. PBMCs were adjusted to the concentrations of more than 1 × 10⁶ cells/L and incubated with various antibodies. Anti-human CD antibodies: CD45-ECD, CD4-FITC, CD3-PC5, CD25-APC, and CD127-PE (Beckman Coulter, USA) were used for Treg detection. Gates were set as high side scatter (SS) and CD45+ cells, followed by CD3+CD4+ cells, CD25+CD127− cells; anti-human CD antibodies: CD45-ECD, CD4-FITC, CD3-PC5 (Beckman Coulter, USA), and IL-17-PE (eBioscience, USA) were used for Th17 detection. Gates were set as high SS and CD45+ cells, followed by CD3+CD4+ cells and CD4+IL-17+ cells. Cells were treated with PerNix-nc kit (Beckman Coulter, USA) before being subjected to Th17 analysis by flow cytometry. All samples were analyzed by Flow Cytometer Navios (Beckman Coulter, USA). The results are expressed as the percentage of Th17 or Treg in PBMCs.

Luo J., Zhang M., Yan B., Zhang K., Chen M, & Deng S. (2016). Imbalance of Th17 and Treg in peripheral blood mononuclear cells of active tuberculosis patients. The Brazilian Journal of Infectious Diseases, 21(2), 155-161.

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