A single-cell suspension was obtained from the spleens of NZB/W and NZW mice at 38-weeks-of age. Briefly, spleens were dissociated using a wire mesh and the cell suspension was centrifuged for 5 minutes at 300 × g. Cells were treated with RBC lysis buffer for 5 minutes at room temperature to lyse erythrocytes, washed 2 times with 1X PBS and then resuspend in flow cytometry staining buffer. Cells were stained with Allophycocyanin (APC)-conjugated CD3, FITC-conjugated CD4, eFluor450 (eF450)-conjugated CD8a, PerCP-CY5.5-conjugated CD25, and PE-conjugated Foxp3, or Fitc-CD4, APC-ROR-γ, and PE-IL-17, or APC-CD3, Fitc-CD4, and PE-CD8 anti-mouse mAbs (eBioscience, San Diego, CA, USA). Fluorescence was measured using a FACS Aria 1 (BD Biosciences, San Jose, CA) and data was analyzed by FlowJo software (Tree Star, Ashland, OR, USA).
Pe conjugated foxp3
Manufactured by Thermo Fisher Scientific
Sourced in United States
PE-conjugated Foxp3 is a laboratory reagent used for the detection and analysis of Foxp3 (Forkhead box P3) protein, a transcription factor that is a critical regulator of regulatory T cell (Treg) development and function. The PE (Phycoerythrin) conjugation allows for the fluorescent labeling and detection of Foxp3-positive cells using flow cytometry or other immunoassay techniques.
Automatically generated - may contain errors
Lab products found in correlation
Apc conjugated cd25, by Thermo Fisher Scientific (2 mentions)
Facsaria 1, by BD (1 mentions)
Efluor 450, by Thermo Fisher Scientific (1 mentions)
Facscanto 2, by BD (1 mentions)
Facscalibur system, by BD (1 mentions)
Fluorescein isothiocyanate fitc conjugated cd4, by Thermo Fisher Scientific (1 mentions)
4 protocols using pe conjugated foxp3
1
Murine Splenic Immune Cell Profiling
2
Multiparametric Flow Cytometry Analysis
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The cellular populations were consequently phosphate-buffered saline, (PBS)-washed, placed into incubation with FITC-labeled anti-CD4 (Solarbio Life Sciences, Beijing, China), and then fluorescently labeled monoclonal antibodies, (mAbs), PE-conjugated RORγt (BD Bioscience, La Jolla, CA, USA) and PE-conjugated Foxp3 (eBioscience, San Diego, CA, USA), in the dark (at 4° C) for 40 min. Then, the cellular populations were assessed on FACS Canto II flow cytometry (BD Bioscience, La Jolla, CA, USA), with data analyzed through FlowJo software.
3
Phenotyping Regulatory T Cells in PBMCs
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Peripheral blood mononuclear cells (PBMCs) were freshly isolated. To analyze intracellular cytokines, FACS was used to detect the CD25 + FoxP3+ cells by gating CD4+ cells, CCR4+ CCR6+ cells, CD4+ CD45RA+ cells and CD4+ CD45RO+ cells in PBSCs using a FACSCalibur system (BD Biosciences, San Jose, CA). FITC conjugated CD4, APC conjugated CD25, PE conjugated Foxp3, PE conjugated CCR4, APC conjugated CCR6, PE conjugated CD45RA, APC conjugated CD45RO, and appropriate isotype controls were purchased from eBioscience (Miltenyi Biotec, CA, USA).
4
Analyzing Th17 and Treg Cells by Flow Cytometry
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To investigate the Th17 and Treg cells, peripheral blood mononuclear cells were first separated and prepared. The percentages of Th17 and Treg cells and the Th17/Treg ratio were ascertained by flow cytometry as previously described (Zhang et al., 2021 (link)). The antibodies used in the experiment, including APC‐Cy7‐conjugated CD3, fluorescein isothiocyanate (FITC)‐conjugated CD4, APC‐conjugated CD25, PE‐conjugated Foxp3, and PE‐conjugated IL‐17, were obtained from eBioscience (San Diego, CA, USA). Data were determined by a flow cytometer (Beckman Coulter, Fullerton, CA, USA).
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