Immature DC, mDC-LPS, mDC-Pam3CSK4 and, iDC treated with PGE2 were harvested by gentle flushing with 1 mL of culture medium, centrifuged for 5 min at 340 g, incubated in staining buffer (PBS, 2 mM EDTA, 0.5% FBS) with a 1:100 dilution of Human TruStain FcX (Biolegend) for 5 min at room temperature. Cells were then stained in cold staining buffer for 25 min with fluorophore-conjugated antibodies (APC-Cy7 labeled anti-HLA-DR (clone: L243, 1/400), PE labeled anti-CD80 (clone: 2D10, 1/200), PerCP-Cy5.5 labeled anti-CD86 (clone: IT2.2, 1/400), Pacific Blue labeled anti-PD-L1 (clone: 29E.2A3, 1/400), PerCP-Cy5.5 labeled anti-CCR5 (clone: J418J1, 1/400), and PE labeled anti-CCR7 (clone: G043H7, 1/400)) from Biolegend. After staining, the cells were washed with cold staining buffer, centrifuged twice for 5 min at 340 g at 4°C, and analyzed by flow cytometry using LSR-II flow cytometer (BD Biosciences) and the associated BD FACSDiva software. Data were then analyzed using the FlowJo 7.6.5 software (TreeStar).
Pe labeled anti ccr7
Manufactured by BioLegend
The PE labeled anti-CCR7 is a laboratory reagent used for the detection and analysis of the CCR7 receptor on cells. It is a fluorescent-labeled antibody that binds to the CCR7 cell surface marker, allowing for its identification and quantification through flow cytometry or other immunoassay techniques.
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Lab products found in correlation
Human trustain fcx, by BioLegend (1 mentions)
Pe labeled anti cd80, by BioLegend (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
Ficoll paque plus gradient, by GE Healthcare (1 mentions)
Human cd4 microbeads, by Miltenyi Biotec (1 mentions)
Anti cd19, by BioLegend (1 mentions)
Anti cd8, by BioLegend (1 mentions)
Facsaria, by BD (1 mentions)
2 protocols using pe labeled anti ccr7
1
Phenotypic characterization of dendritic cell subsets
2
Flow Cytometric Profiling of CD4+ T Cell Subsets
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Peripheral blood mononuclear cells (PBMCs) were isolated from blood by density-gradient centrifugation using Ficoll-Paque Plus gradient (GE Healthcare). CD4+ T cells were enriched by positive selection using human CD4 MicroBeads (Miltenyi Biotec), and then CD4+ T cell subsets were sorted to 99% purity with a FACSAria (BD). For in vitro TCR stimulation, CD4+ naive and CXCR5+ cells were stimulated with anti-CD3 (5 µg/ml) and anti-CD28 (1 µg/ml) mAbs. PBMCs were stained with the following antibodies: PE-Texas Red–labeled anti-CD4 (clone: MHCD0417; Life Technologies), QD655-labeled anti-CD45RA (clone: MEM-56; Life Technologies), PE-labeled anti-CCR7 (clone: G043H7; BioLegend), purified mouse monoclonal IgG2b anti-CXCR5 (BRL-1, clone: 51505; R&D Systems) revealed by FITC-labeled anti–mouse IgG2b (Southern Biotech), PE-cyanine 5 (PeCy5)–labeled anti-CD183 (CXCR3; clone: 1C6/CXCR3; BD), PE-labeled anti-CD196 (CCR6; clone: 11A9; BD), BV785-labeled anti-CD279 (PD-1; clone: EH12.2H7; BioLegend). PeCy5-labeled anti-CD56 (clone: A07789; Beckman Coulter), anti-CD14 (clone: A07765; Beckman Coulter), anti-CD19 (clone: HIB19; BioLegend), anti-CD25 (clone: BC96; BioLegend), and anti-CD8 (clone: HIT8a; BioLegend) were included as a dump channel to exclude contaminant cells.
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