The antibodies (BD Biosciences, San Jose, CA) used for flow cytometry were: CD4-PE (RM4-5), CD4-v450 (RM4-5), CD8-APC (53-6.7), CD69-FITC (H1.2F3), CD134-Biotin (OX-86), CD62L-APC (MEL-14), B220-APC (RA3-6B2), CD86-PE (GL1), CD22-PE (Cy34.1), CD25-APC.Cy7 (PC61), and FoxP3-PE (150D). Biotinylated antibodies were detected using FITC-conjugated streptavidin (BD Biosciences). Flow cytometric analysis was performed using various combinations of these antibodies on single cell suspensions of splenocytes. For intracellular FoxP3 staining, surface-stained cells were treated with fixation/permeabilization buffer and stained with FoxP3-PE (150D) using the BioLegend FoxP3 flow kit, following the manufacturer’s protocol (BioLegend). Stained cells were analyzed in the UNMC Flow Cytometry Research Facility using the BD LSR II flow cytometer. Data were analyzed using FACSDiva software, version 6.1.2 (BD Biosciences). For analysis of T regulatory cells, splenocytes were isolated from mice that were 3–4 months of age. For all other flow cytometry analyses, splenocytes were collected from mice that were 6–12 months of age.
Foxp3 pe 150d
Manufactured by BioLegend
FoxP3-PE (150D) is a fluorochrome-conjugated antibody that specifically binds to the FoxP3 transcription factor. FoxP3 is a key regulator of the development and function of regulatory T cells (Tregs). The FoxP3-PE (150D) antibody can be used to identify and analyze Tregs in various cell samples.
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Lab products found in correlation
Facsdiva software, by BD (1 mentions)
Cd69 fitc, by BD (1 mentions)
Cd86 pe, by BD (1 mentions)
Foxp3 pe, by BD (1 mentions)
B220 apc, by BD (1 mentions)
Fitc conjugated streptavidin, by BD (1 mentions)
Cd62l apc, by BD (1 mentions)
Cd22 pe, by BD (1 mentions)
Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions)
Facsymphony a5, by BD (1 mentions)
Brilliant stain buffer, by BD (1 mentions)
Cd3 bv785 okt3, by BioLegend (1 mentions)
Dnase 1, by STEMCELL (1 mentions)
Cd25 apc 2a3, by BD (1 mentions)
2 protocols using foxp3 pe 150d
1
Multicolor Flow Cytometry for T Cell Analysis
2
Multiparametric Phenotyping of Immune Cells
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At the day of the experiment, cryopreserved samples were thawed in RPMI media supplemented with 30% FBS, 10ug/mL DNAse I (Stemcell Technologies). Cells were rested for 20 min in 37°C, spun and stained on ice with LIVE/DEAD™ Fixable Violet, washed with PBS 2% FBS and stained for surface markers with CD3 BV785 [OKT3] (Biolegend), CD4 APC-R700 [RPA-T4] (BD), CD8 PE-Cy7 [SK1] (BioLegend), CD25 APC [2A3] (BD), CD69 BUV395 [FN50] (BD), CD1a BUV496 [HI149] (BD), CD14 PB [63D3] (BioLegend) antibodies in PBS supplemented with 1% BSA and brilliant stain buffer (BD) for 30 min on ice. Cells were washed, fixed and permeabilized using Foxp3/Transcription Factor Staining Buffer Set (Invitrogen) and stained with FOXP3 A488 [259D] (BioLegend) and FOXP3 PE [150D] (BioLegend) antibodies or isotype controls 488 and PE [MOPC-21] (BioLegend). Cells were washed and analyzed using BD FACSymphony™ A5.
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