Leukocytes were isolated from serially collected peripheral blood samples, spleens or liver grafts. Splenocytes were isolated using standard protocol. To obtain single cell-suspensions from cardiac or liver grafts, tissue was cut into small pieces (2 mm) and digested with collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ). The resulting suspension was run through a 70 μm filter and washed with PBS. After centrifugation, the leukocytes were purified using lymphocyte separation medium (Fisher Scientific, Pittsburgh, PA). Phenotypical analysis was performed by using several panels of fluorescein-labeled anti-mouse mABs (against CD3, CD4, CD8, Ly6G, Ly6C, MHCII, CCR2, CXCR1, CD80, CD86) or rat monoclonal antibodies (CD3, CD4, or CD8). All antibodies were purchased from BD Biosciences. Frequencies of T-regulatory cells were identified using APC conjugated CD4, FITC conjugated-CD25, and PE conjugated-Foxp3 (Biolegend) monoclonal antibodies according to the manufacture instructions. Staining was performed with antibodies above (1 μg/106 cells) at 4 °C for 30 min, washed, and analyzed by FACS (BD Biosciences).
Pe conjugated foxp3
Manufactured by BioLegend
Sourced in United States
PE conjugated-Foxp3 is a laboratory reagent used for the detection and analysis of Foxp3-expressing cells, such as regulatory T cells, through flow cytometry. Foxp3 is a transcription factor that is essential for the development and function of regulatory T cells. The PE (phycoerythrin) fluorescent dye is conjugated to the Foxp3 antibody, allowing for the identification and quantification of Foxp3-positive cells in a sample.
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Lab products found in correlation
Lymphocyte separation medium, by Thermo Fisher Scientific (1 mentions)
Fitc conjugated cd25, by BioLegend (1 mentions)
Collagenase 4, by Worthington (1 mentions)
Fluorescence activated cell sorting (facs), by BD (1 mentions)
Foxp3 staining set, by Thermo Fisher Scientific (1 mentions)
Pe conjugated ki 67, by BioLegend (1 mentions)
Alexa fluor 488, by BioLegend (1 mentions)
3 protocols using pe conjugated foxp3
1
Isolation and Analysis of Leukocytes
2
Immunophenotyping of Fixed PBMCs
3
Quantifying Tumor-Infiltrating Tregs in Mice
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Single cell suspensions were prepared from the tumor tissue of mice. Cells were stained with the following mouse-specific antibodies: FITC-conjugated CD4, APC-conjugated CD25, and PE-conjugated Foxp3 (cat Nos. 100406, 100910, and 320008, respectively; Biolegend Inc.). Analysis of cell surface markers was performed using FlowJo (FlowJo LLC) and gated according to surface markers and negative controls. Isotype-matched IgG staining was used as the negative control (Thermo Fisher Scientific, Inc.). Tregs were isolated from the tumor tissue of mice as follow single cell suspensions were made from the tumor tissues. These cells were analysed with fluorescence-conjugated antibodies and isotype-matched IgG controls. The cells were analysed on an FACScan flow cytometer.
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