The ex vivo detection of human IL-17+ T cells was performed as previously described (de Beaucoudrey et al., 2008 (link)). In brief, PBMCs were purified on Ficoll-Paque PLUS (GE Healthcare) and resuspended in RPMI + 10% FBS (Invitrogen). We took aliquots of 106 nonadherent cells/ml and either left them unstimulated or stimulated them with 40 ng/ml PMA (Sigma-Aldrich) + 10−5 M ionomycin (Sigma-Aldrich) for 12 h. All cells were treated with 1 µl/ml GolgiPlug for the final 12 h of culture. Surface staining was performed with PE-Cy5–conjugated anti–human CD3 (PE Biosciences) Ab, and intracellular staining was performed with Alexa Fluor 488–conjugated anti–human IL-17 (eBioscience) and PE-conjugated anti–human IFN-γ (BD) or PE-conjugated anti–human IL-22 (R&D Systems) Abs. Cells were analyzed with a FACScan machine and CellQuest software (both from BD).
Pe conjugated anti human ifn γ
Manufactured by BD
Sourced in United Kingdom
PE-conjugated anti–human IFN-γ is a laboratory reagent used for the detection and quantification of human interferon-gamma (IFN-γ) in various biological samples. It is a fluorescently labeled antibody that specifically binds to IFN-γ, allowing its identification and measurement using flow cytometry or other immunoassay techniques.
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Lab products found in correlation
Ficoll paque plus, by GE Healthcare (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Cellquest software, by BD (1 mentions)
Facscan machine, by BD (1 mentions)
Il 10, by BD (1 mentions)
Granzyme b, by BD (1 mentions)
Intrastain, by Agilent Technologies (1 mentions)
Brefeldin a, by Merck Group (1 mentions)
Tnf α, by BD (1 mentions)
Anti cd28, by BD (1 mentions)
Interleukin 2 (il 2), by BD (1 mentions)
2 protocols using pe conjugated anti human ifn γ
1
Detecting Human IL-17+ T Cells
2
Cytokine Profiling of CMV-CTL Effector Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess the effector activity of CMV-CTL by intracellular cytokine staining, expanded cells were co-cultured for 6 h with CMV-loaded or control feeders labelled with CFSE at a 2.5:1 ratio, in the same way as for surface activation marker expression analysis. The incubation was done in the presence of 1 μg/ml anti-CD28 (BD Bioscience) and 1 μg/ml of brefeldin A (Sigma-Aldrich). Following the incubation, 1 × 106 cells were stained with either PE-conjugated anti-human IFN-γ, IL-2, IL-10, TNF-α, or Granzyme B (BD Biosciences) and with CD8-PerCP and CD4-APC-Cy7 monoclonal antibodies, then fixed and permeabilized (Intrastain; DakoCytomation, Ely, UK), according to the manufacturer’s instructions.
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