All mAbs were purchased from BD PharMingen (San Diego, CA), except for anti-CCR2 (R&D Systems, Minneapolis, MN). Mouse Treg cell staining kit was purchased from eBioscience (San Diego, CA). The appropriate isotype-matched irrelevant Abs served as controls. For CFSE labeling, T cells (1×107 cells/mL) were incubated with 2 µM CFSE (Molecular Probes, Eugene, OR) for 10 minutes. For intracellular staining (Foxp3), cells were first permeabilized with 0.1% saponinb. Flow analyses were performed on a FACSCalibur flow cytometer (BD Biosciences) and the data was analyzed by FlowJo software.
Mouse treg cell staining kit
Manufactured by Thermo Fisher Scientific
The Mouse Treg cell staining kit is a laboratory reagent designed to identify and quantify regulatory T cells (Tregs) in mouse samples. The kit contains antibodies and buffers necessary for the staining and flow cytometric analysis of Tregs.
Automatically generated - may contain errors
3 protocols using mouse treg cell staining kit
1
Regulatory T Cell Isolation and Characterization
2
Multiparameter Immune Cell Profiling
3
Assessing T Reg Cell Modulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The effects of hyperforin and HPE on T reg cells were assessed by flow cytometry, using a mouse T reg cell staining kit (eBioscience, San Diego, CA). Spleen cells were isolated from all mice as outlined above. For marker staining, aliquots of 10 6 cells were washed and stained with fluorescein isothiocyanate (FITC)-conjugated rat anti-mouse CD4 (clone RM4) and phycoerythrin (PE)-conjugated rat anti-mouse CD25 (clone PC61.5) antibodies at 4 C for 30 min. To stain for FoxP3, these cells were then fixed and permeabilized with kit-provided fixation/permeabilization buffer. The cells were then washed with permeabilization buffer and incubated in this buffer containing PE-Cy5 rat anti-mouse Foxp3 (clone FJK16s) antibody or isotype control (PE-Cy5 mouse IgG 2a isotype control) at 4 C for 30 min. Ultimately, all samples were washed and suspended in kit-provided staining buffer and then analyzed in a FACSCalibur system (BD Bioscience, San Diego, CA) with associated WinMDI 2.9 software. A minimum of 20 000 events in the lymphocyte gate was collected per sample.
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