The ability of the study's assay to detect intracellular cytokines in T cells and monocytes in whole blood samples was assessed with particular interest focused on the following cytokines, all PE-conjugated: TNF-α (BD FastImmune, Clone 6401.111), IFN-γ (BD FastImmune, Clone 25723.11), IL-10 (BD Pharmingen, Clone JES3-9D7), IL-6 (BD Pharmingen, Clone MQ2-13A5), IL-4 (BD FastImmune, Clone 3010.211), IL-2 (BD Pharmingen, Clone MQ1-17H12), TGF-β (BD Pharmingen, Clone TW4-2F8), IL-12 p40 (BD Pharmingen, Clone C11.5) and IL-12 p70 (BD Pharmingen, Clone 20C2). Stimulation was performed as described earlier and the antibody labelling was performed using 50 μl of whole blood as already described with anti-CD3-PerCP used at a ratio of 1:25, anti-CD14-APC at a ratio of 1:50 and anti-cytokine-PE at a ratio of 1:12.5. One tube was included with unstimulated blood and labelled with anti-TNF-α−PE at the same ratio to account for any background noise and another tube was included with stimulated blood but labelled with an isotype control at the same ratio.
Clone jes3 9d7
Manufactured by BD
The Clone JES3-9D7 is a laboratory equipment designed for specific experimental functions. It serves as a core component for scientific investigations, providing essential functionalities for researchers. The detailed specifications and intended use of this product are not available within the scope of this response.
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Lab products found in correlation
Clone mq1 17h12, by BD (1 mentions)
Mp4 25d2, by BD (1 mentions)
Fix perm, by BD (1 mentions)
Clone 4s b3, by BD (1 mentions)
Clone mab11, by BD (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Fcr blocking reagent, by Miltenyi Biotec (1 mentions)
Diva software, by BD (1 mentions)
Cytofix cytoperm kit, by BD (1 mentions)
Perm wash, by BD (1 mentions)
2 protocols using clone jes3 9d7
1
Intracellular Cytokine Detection in Whole Blood
2
Multicolor Flow Cytometry Analysis of Immune Cell Subsets
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PBMCs were washed with FACS buffer and stained for cell viability with the fixable live/dead Aqua staining kit (Molecular Probes) in the presence of Fc-R blocking reagent (Miltenyi biotech), and the antibodies against the surface markers anti-CD3-APC, CD45-V450 and anti-CD11b-APC/Cy7 indicated above. Surface staining was carried out for 25 min at 4°C. Cells were washed, and intracellular staining was carried out with BD cytofix-cytoperm kit (BD Biosciences). Cells were fixed with Fix-Perm (BD Biosciences) for 20 min at room temperature and permeabilized with Perm-Wash (BD Biosciences) for 30 min at room temperature. Cells were stained with two combinations of antibodies diluted in Perm-Wash: anti-IFN-γ (PE, 1:100, clone 4S.B3, BD Biosciences), anti-IL-4 (FITC, 1:100, clone MP4-25D2, BD Biosciences), and anti-IL-17A (PE/Cy7, 1:100, clone BL168, BioLegend) or anti-TNF-α (AlexaFluor-488, 1:100, clone MAb11, BD Biosciences) and anti-IL-10 (PE, 1:30, clone JES3-9D7, BD Biosciences). Data were acquired on a FACSCanto II flow cytometer (BD Biosciences) with Diva software (BD Biosciences) and analyzed with FlowJo (v10, BD Biosciences).
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