For analysis of surface markers, a single-cell suspension from mouse spleens was stained with anti-CD4-PE/Cy7 antibody (BioLegend, 100528) or anti-CD25-APC antibody (BioLegend, 102012) for 30 min on ice in the dark. After fixation and permeabilization with a True-Nuclear Transcription Factor Buffer Set (BioLegend, 424401), cells were stained with anti-Foxp3-PE antibody (eBioscience, 126404) for detection of Tregs. For intracellular cytokine measurement, cells were incubated with Cell Activation Cocktail (BioLegend, 423303) at 37 °C in the dark for 6 h. Anti-IL-17-PE antibody (eBioscience, 12-7177-81) was used to determine intracellular expression of IL-17. The percentages of positive cells were determined with a flow cytometer (Cytomics FC 500; Beckman-Coulter) and analyzed using FlowJo_V10 software.
Anti cd25 apc antibody
Manufactured by BioLegend
The Anti-CD25-APC antibody is a fluorescently-labeled monoclonal antibody that binds to the CD25 receptor, also known as the interleukin-2 receptor alpha chain (IL-2Rα). The APC (Allophycocyanin) fluorescent dye is conjugated to the antibody, allowing for detection and analysis of CD25-expressing cells using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Cell activation cocktail, by BioLegend (1 mentions)
True nuclear transcription factor buffer set, by BioLegend (1 mentions)
Pe anti foxp3 antibody, by Thermo Fisher Scientific (1 mentions)
Cytomics fc500, by Beckman Coulter (1 mentions)
Red blood cell lysis solution, by Miltenyi Biotec (1 mentions)
Cd4 t cell isolation kit 2, by Miltenyi Biotec (1 mentions)
Macs buffer, by Miltenyi Biotec (1 mentions)
Facsaria cell sorter, by BD (1 mentions)
Macs ls column, by Miltenyi Biotec (1 mentions)
2 protocols using anti cd25 apc antibody
1
Mouse Splenocyte Phenotyping by Flow Cytometry
2
Isolation and Transfer of Murine CD4+ T Cells
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Spleen cells were removed from BALB/c mice and suspended into magnetic-activated cell separation (MACS) buffer (Miltenyi Biotec, Bergisch Gladbach, Germany). Erythrocytes were removed by a red blood cell lysis solution (Miltenyi Biotec). CD4 + T cells were isolated by a CD4 + T cell isolation kit II (Miltenyi Biotec), MidiMACS Separator LS (Miltenyi Biotec), and MACS LS column (Miltenyi Biotec). 22 The resulting population was .92% CD4 + T cells. CD4 T cells were labeled with anti-CD45RB FITC antibody (BioLegend, San Diego, CA), anti-CD4 PE antibody (BioLegend), and anti-CD25 APC antibody (BioLegend), and then CD4 + CD25 2 CD45RB high (defined as 20% brightest staining) 23 was sorted by a BD FACSAria cell sorter. C.B-17/SCID mice were injected intraperitoneally with 100 mL of phosphate-buffered saline (PBS) containing 2.0 • 10 5 CD4 + CD25 2 CD45RB high T cells. 22
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