HVS-T cells from six healthy donors and P were cultured in the presence of IL-2 at 20 IU/ml (Roche). HVS-T cells were harvested, counted, and topped up to 106 cells/ml without recombinant IL-2. 200 µl cells per well were plated to 96-well round bottom tissue-culture plates. After 24 h culture, cells were stimulated with 25 ng/ml PMA and 0.5 µM Ionomycin for 3 h. Protein transport inhibitor (eBioscience) was added to each well. After another 3 h, cells were harvested for surface staining with FcBlock, anti-CD271-FITC Abs, and Zombie-NIR live-dead exclusion dye (BioLegend). Cells were then fixed, permeabilized with FOXP3/perm kit (Thermo Fisher Scientific), and subjected to overnight ICS with anti-GATA3-BV421 (BD Biosciences), anti-IL-13-BV711 (BD Biosciences), anti-IFN-γ-eFluor450 (eBioscience), anti-IL-9-PERCP/Cy5.5 (BioLegend), anti-IL-5-PE (BioLegend), anti-T-bet-PE/Cy7 (BioLegend), anti-IL-10-PE/Dazzle594 (BioLegend), anti-IL-22-APC/Fire750 (BioLegend), anti-TNF-BV510 (BioLegend), anti-RORγ/RORγT-BV650 (BD Bioscience), anti-IL-17A-BV785 (BioLegend), and anti-IL-4-Alexa 647 (BioLegend), followed by flow cytometry analysis with an Aurora cytometer (Cytek).
Anti tnf bv510
Manufactured by BioLegend
Anti-TNF-BV510 is a fluorophore-conjugated antibody product manufactured by BioLegend. It is designed for the detection and analysis of tumor necrosis factor (TNF) in various biological samples using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Protein transport inhibitor, by Thermo Fisher Scientific (1 mentions)
Aurora cytometer, by Cytek (1 mentions)
Anti ifnγ efluor 450, by Thermo Fisher Scientific (1 mentions)
Fc block, by BioLegend (1 mentions)
Anti gata3 bv421, by BD (1 mentions)
Anti t bet pe cy7, by BioLegend (1 mentions)
Zombie nir live dead exclusion dye, by BioLegend (1 mentions)
Anti il 5 pe, by BioLegend (1 mentions)
Interleukin 2 (il 2), by Roche (1 mentions)
Anti ifn γ apc, by BioLegend (1 mentions)
Anti il 2 bv510, by BioLegend (1 mentions)
Anti il2 pe cy7, by BioLegend (1 mentions)
Anti il 2 pacificblue, by BioLegend (1 mentions)
Lsrfortessa, by BD (1 mentions)
Facscanto 2, by BD (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Golgistop, by BD (1 mentions)
Anti ifn γ fitc, by BioLegend (1 mentions)
2 protocols using anti tnf bv510
1
Profiling Cytokine Expression in Stimulated HVS-T Cells
2
Multiparameter Flow Cytometry Analysis
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Cells were stimulated (3 h) with phorbol-12-myristate 13-acetate (PMA; 50 ng ml−1, Sigma-Aldrich, P1585), ionomycin (1 μg ml−1, Sigma-Aldrich, I3909) and Golgi Stop (1:100, BD Biosciences, 554724). The cells were then fixed and permeabilized (1:100, BD Biosciences, 554714) and stained with anti-IFNγ–APC (1:100, BioLegend, 505810), anti-IFNγ–FITC (1:100, BioLegend, 505806), anti-TNF–BV510 (1:100, BioLegend, 506339), anti-TNF–BV5711 (1:100, BioLegend, 506349), anti-TNF–PE (1:100, BioLegend, 506306), anti-IL2–Pecy7 (1:100, BioLegend, 503832), anti-IL-2–Pacific Blue (1:100, BioLegend, 503820), anti-IL-2–BV510 (1:100, BioLegend, 503833), and analysed using a LSRFortessa or FACSCanto II (Becton Dickinson).
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