After thawing PBMCs in PBS containing the endonuclease benzonase (Novagen/Merck Milipore Schwalbach, Germany), cells were directly stained in PBEA buffer (PBS, 0.5% BSA, 2 mM EDTA and 0.01% NaN3 ) for 30 min on 4°C. An overview of the B cell-, T cell- and NK cell/regulatory T cell (Treg)-phenotyping panels including fluorochromes is provided in Table S2. Cells were washed with PBEA buffer and fixed in PBS containing 1% formaldehyde. The following monoclonal antibodies were used after initial testing and titration: anti-CD3 Alexa-eFluor 780, anti-CD4 PE-Cy7, anti-CD19 Pe-Cy7, anti-CD27 APC, anti-CD28 FITC, and anti-CCR7 PE (eBioscience, SanDiego, USA), anti-CD4 PerCP-Cy5.5, anti-CD8+ PerCP-Cy5.5, anti-CD16 PerCP-Cy5.5, anti-CD20 FITC, anti-CD25 V450, anti-CD38 PerCP-Cy5.5, anti-CD45RA HV450, anti-CD69 FITC, anti-CD86 V450, anti-IFN-g APC, anti-IgD PE, anti-TNF-a FITC (Becton Dickenson GmbH, Heidelberg, Germany), anti-IL-2 PacificBlue (BioLegend,); anti-CD56 PE (Miltenyi, Bergisch Gladbach, Germany).
Anti il 2 pacificblue
Manufactured by BioLegend
Sourced in Germany
Anti-IL-2 PacificBlue is a monoclonal antibody that binds to the human interleukin-2 (IL-2) cytokine. It is useful for the detection and quantification of IL-2 in various applications, such as flow cytometry, ELISA, and other immunoassays.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd4 pe cy7, by Thermo Fisher Scientific (1 mentions)
Anti cd28 fitc, by Thermo Fisher Scientific (1 mentions)
Anti cd27 apc, by Thermo Fisher Scientific (1 mentions)
Anti ccr7 pe, by Thermo Fisher Scientific (1 mentions)
Anti cd56 pe, by Miltenyi Biotec (1 mentions)
Anti cd19 pecy7, by Thermo Fisher Scientific (1 mentions)
Anti ifn γ apc, by BioLegend (1 mentions)
Anti il 2 bv510, by BioLegend (1 mentions)
Anti tnf bv510, by BioLegend (1 mentions)
Anti il2 pe cy7, 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 il 2 pacificblue
1
Phenotyping of Immune Cell Subsets
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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