Lamina propria immune cells (2.0×105/200 μl) were stained with fluorophore-conjugated anti-mouse monoclonal antibodies (BD Bioscience), directed against the following cell surface proteins: CD3-PE and CD4-PE-Cy7, in staining buffer (BD Bioscience) and incubated for 30 min at room temperature in the dark. After surface staining, cells were washed in PBS, fixed and permeabilized with FoxP3 staining buffer Kit (BD Bioscience) according to manufacturer’s instructions and incubated with FoxP3-Alexa Fluor 488 and RORγt-Alexa Fluor 647 for 1h at room temperature. Cells were washed twice in FACS Buffer and were analyzed by Accuri c6 flow cytometer (BD Biosciences) with BD Accuri C6 Software (Becton Dickinson). Immune cells were defined by using surface markers: regulatory T cells (Treg) CD3+CD4+FoxP3+; Th17 cells CD3+CD4+RORγt+.
Fluorophore conjugated anti mouse monoclonal antibodies
Manufactured by BD
Fluorophore-conjugated anti-mouse monoclonal antibodies are laboratory reagents designed for the detection and analysis of target proteins or molecules in various experimental applications. These antibodies are conjugated with fluorescent dyes, enabling visualization and quantification of the target analytes through fluorescence-based techniques.
Automatically generated - may contain errors
2 protocols using fluorophore conjugated anti mouse monoclonal antibodies
1
Flow Cytometry Analysis of Treg and Th17 Cells
2
Flow Cytometry Analysis of Treg and Th17 Cells
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Lamina propria immune cells (2.0×105/200 μl) were stained with fluorophore-conjugated anti-mouse monoclonal antibodies (BD Bioscience), directed against the following cell surface proteins: CD3-PE and CD4-PE-Cy7, in staining buffer (BD Bioscience) and incubated for 30 min at room temperature in the dark. After surface staining, cells were washed in PBS, fixed and permeabilized with FoxP3 staining buffer Kit (BD Bioscience) according to manufacturer’s instructions and incubated with FoxP3-Alexa Fluor 488 and RORγt-Alexa Fluor 647 for 1h at room temperature. Cells were washed twice in FACS Buffer and were analyzed by Accuri c6 flow cytometer (BD Biosciences) with BD Accuri C6 Software (Becton Dickinson). Immune cells were defined by using surface markers: regulatory T cells (Treg) CD3+CD4+FoxP3+; Th17 cells CD3+CD4+RORγt+.
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