Unstimulated BAL cells were stained ex vivo with anti-CD3-PE-Cy7 and CCR5-PE (both from BD) and with CD4-PE-Cy5.5 and CD8-Qdot705 (both from Invitrogen). Freshly stimulated BAL cells and stimulated cryopreserved blood cells were washed and stained with anti-CD4-PE-Cy5.5 and CD8-Qdot705 (both from BD), permeablized, and stained intracellularly with CD3-APC-H7, IFN-γ-Alexa700, and interleukin 2 (IL-2)-APC (all from BD) and with tumor necrosis factor α (TNF-α)-PE-Cy7 (eBiosciences). Cells were acquired on a BD Fortessa, using FACSDiva software, and data were analyzed using FlowJo (TreeStar) and Pestle and Spice [25 (link)]. A positive cytokine response was defined as a level that was twice the background level, a net response of >0.05%, and an event cutoff of 10 events, and all data are reported after subtraction of the background level.
Cd8 qdot705
Manufactured by Thermo Fisher Scientific
The CD8-Qdot705 is a fluorescent conjugate used for cell labeling and flow cytometry analysis. It consists of the CD8 antibody coupled to a Quantum Dot 705 fluorescent label.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd3 pe cy7, by BD (1 mentions)
Ccr5 pe, by BD (1 mentions)
Cd3 apc h7, by BD (1 mentions)
Cd4 pe cy5, by Thermo Fisher Scientific (1 mentions)
Ifn γ alexa700, by BD (1 mentions)
Tnf α pe cy7, by Thermo Fisher Scientific (1 mentions)
Cd19 apc cy7, by BD (1 mentions)
Cd57 pacific blue, by BioLegend (1 mentions)
Cd56 pe cy7, by BD (1 mentions)
Facsaria 2, by BD (1 mentions)
2 protocols using cd8 qdot705
1
Ex vivo Analysis of Cytokine Responses
2
Multiparametric Cell Sorting Protocol
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Cells were stained with fluorochrome-conjugated antibodies and incubated for 30 min on ice. The following antibodies were used for labeling cells before sorting: CD56 PE-Cy7 (BD), CD3 PE-Cy5.5 (Invitrogen), CD19 APC-Cy7 (BD), CD8 QDot 705 (Invitrogen), CD45 QDot 655 (Invitrogen), and CD57 Pacific Blue (BioLegend). Cells were sorted with a FACSAria II (BD).
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