Cells were stained with PE-conjugated anti-PD-L1, PerCp-conjugated anti-7-AAD, APC-Cy7-conjugated anti-CD3, PE-Cy7-conjugated anti-CD8, APC-conjugated anti-IFN-γ, and FITC-conjugated anti-granzyme B antibodies (BioLegend, USA). Dead cells were stained using 7-AAD. Among them, IFN-γ and granzyme B were used for intracellular staining as follows: cells were first fixed with 2% paraformaldehyde and permeabilized with 0.1% saponin in phosphate buffered saline (PBS) buffer. Next, cells were incubated in the dark for 15 min on ice with antibodies labeled with fluorochrome. For surface assessment, cells were incubated with fluorochrome-labeled antibodies directly. The cell phenotype was determined using cytofluorimetric analysis by flow cytometer (BD FACSCanto II, USA).
Pe cy7 conjugated anti cd8
Manufactured by BioLegend
Sourced in United States
PE-Cy7-conjugated anti-CD8 is a fluorescently-labeled antibody that binds to the CD8 surface antigen. The PE-Cy7 fluorophore is attached to the anti-CD8 antibody, allowing for the detection and identification of CD8-positive cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Apc cy7 conjugated anti cd3, by BioLegend (2 mentions)
Facscanto 2, by BD (2 mentions)
Apc conjugated anti ifn γ, by BioLegend (1 mentions)
Pe conjugated anti pd l1, by BioLegend (1 mentions)
Fitc conjugated anti cd3, by BioLegend (1 mentions)
Pe cy5 conjugated anti cd4, by BioLegend (1 mentions)
Fc500, by Beckman Coulter (1 mentions)
Fixation permeabilization solution kit, by BD (1 mentions)
Fitc conjugated anti ifn γ, by BD (1 mentions)
Brefeldin a, by Merck Group (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Pe conjugated anti ifn γ, by BioLegend (1 mentions)
Apc cy7 conjugated anti cd14, by BioLegend (1 mentions)
Peptivator, by Miltenyi Biotec (1 mentions)
Apc cy7 conjugated anti cd19, by BioLegend (1 mentions)
Lsrfortessa, by BD (1 mentions)
Cytofix cytoperm, by BD (1 mentions)
Golgistop, by BD (1 mentions)
Golgiplug, by BD (1 mentions)
4 protocols using pe cy7 conjugated anti cd8
1
Multiparametric Flow Cytometry Analysis
2
Murine Spleen Lymphocyte Profiling
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Spleens of the control group or LPS-challenged apoM+/+ and apoM−/− mice were harvested aseptically and filtered through cell strainers to produce a single-cell suspension. After lysis of red blood cells, the spleen suspensions were washed three times with PBS and adjusted to a concentration of 1.0 × 107/mL. To detect the changes in lymphocyte subgroups, the prepared splenocytes (1.0 × 107/mL, 100 μL) were incubated with 0.5 μL each of fluorescently labeled antibodies for 30 minutes at 4°C protected from light and washed three times with PBS before analysis. Fluorescently labeled antibodies included FITC-conjugated anti-CD3, PE-Cy5 conjugated anti-CD4, and PE-Cy7 conjugated anti-CD8 (BioLegend, San Diego, CA). The stained cells were then analyzed by flow cytometry immediately (FC500, Beckman, USA).
3
Cytokine Production in Activated T Cells
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DCs were stimulated with HIV‐1 Cap‐RNA58, R848 or a combination, for 48 h and subsequently cocultured with allogeneic PBLs in IMDM complete, using a 1:8 ratio. As a positive control, T cells were stimulated with anti‐CD3 (1:10,000; Sanquin) and anti‐CD28 (1 μg/ml; Sanquin). After 3 days, IL‐2 was added to the cocultures. After 6 days, cells were restimulated using PMA (10 ng/ml; Sigma) and ionomycin (1 mg/ml; Sigma) for 6 h, and brefeldin A (10 μg/ml; Sigma) for the final 4 h, followed by fixation and permeabilization using the fixation/permeabilization solution kit (BD Biosciences) according to manufacturer's instructions. Cells were then stained using APC‐Cy7‐conjugated anti‐CD3 (1:100, 300317; Biolegend), PE‐Cy7‐conjugated anti‐CD8 (1:100, 344711; Biolegend), FITC‐conjugated anti‐IFNγ (1:5, 340449; BD Biosciences), PE‐conjugated anti‐Perforin (1:10, 12‐9994‐42; Thermo Fisher) and AF700‐conjugated anti‐Granzyme B (1:200, 560213; BD Biosciences) and flow cytometry was performed using FACS Canto II (BD Biosciences) and analyzed with FlowJo software v10.7.
4
SARS-CoV-2 Spike Protein T-Cell Assay
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Overnight-rested PBMCs were stimulated with SARS-CoV-2 overlapping peptide pools against SARS-CoV-2 spike protein (PepTivator, Miltenyi Biotec) at a final concentration of 1 μg ml -1 for 1 h in the presence of 1 μg ml -1 monoclonal antibodies CD28 and CD49d, and then for an additional 5 h with GolgiPlug and GolgiStop (BD Biosciences). Dead cells were labelled using Fixable Viability eFluor 780 dye (Thermo Fisher Scientific). Surface markers were stained using BV786-conjugated anti-CD3 (BD Biosciences, 565491; diluted 1:100), BUV486-conjugated anti-CD4 (BD Biosciences, 612937; 1:50), PE-Cy7-conjugated anti-CD8 (BioLegend, 301012; 1:100), APC-Cy7-conjugated anti-CD14 (BioLegend, 301820; 1:100), APC-Cy7-conjugated anti-CD56 (BioLegend, 362512; 1:100) and APC-Cy7-conjugated anti-CD19 (BioLegend, 302218; 1:100). Cells were then washed, fixed with Cytofix/Cytoperm (BD Biosciences) and stained with PE-conjugated anti-IFNγ (BioLegend). Negative controls without peptide stimulation were run for each sample. All results were acquired on a BD LSRFortessa (BD Biosciences) flow cytometer using the BD FACSDIVA v.8.01 software and analysed using FlowJo v.10.6.1 software.
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