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2 protocols using il 2 fitc clone jes6 5h4

1

Comprehensive Immune Cell Profiling

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The following Abs were used for cell surface staining: anti-CD3 Pacific Blue (clone 17A2), anti-CD8 PE Texas Red (clone 53.6.7), anti-CD44 PE-Cy7 (clone IM7), anti-B220 PE Texas Red (clone RA3-6B2), anti-NK1.1 FITC (clone PK136), anti-CD69 Alexa Fluor 647 (clone H1-2F3), anti-Ly6C APC-Cy7 (AL-21), anti-Ly6G PE-Cy7 (clone 1A8) (Biolegend, San Diego, CA, USA), anti-CD4 APC e-Fluor 780 (clone GK1.5), anti-CD62L PE-Cy5 (clone MEL-14) (eBiosciences, San Diego, CA, USA), anti-CD45RB PE (clone 16A), anti-CD3 Alexa Fluor 647 (clone 17A2), anti-CD4 PerCP (clone RM4-5), anti-CD8 PE (clone 53-6.7), anti-CD45 PE-Cy5 (clone 30-F11), anti-CD45 PE (clone 30-F11), anti-CD11c FITC (clone HL3) and anti-CD11b Alexa Fluor 700 (clone M1/70) (BD Biosciences, San José, CA, USA). The abs for intracellular staining included anti-FoxP3 Alexa Fluor 488 (clone MF23), anti-IFNγ Alexa Fluor 700 (clone XMG1.2), TNFα PE-Cy7 (clone MP6-XT22), IL-2 FITC (clone JES6-5H4) (BD Biosciences) and anti-CTLA-4 PE (clone UC10-4F10-11) (Biolegend). A LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (Invitrogen Molecular Probes, Eugene, OR) was used for dead cell exclusion.
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2

Multiparametric Flow Cytometry Analysis

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The Abs used for cell-surface staining included: anti-CD3 Pacific Blue (clone 17A2), anti-CD8 PE Dazzle 594 (clone 53.6.7), anti-CD45 PE-Cy5 (clone 30-F11), anti-Ly-6G PE-Cy7 (clone 1A8), anti-Ly-6C APC-Cy7 (clone AL-21), anti-PD-L1 PE (clone 10F.9G2), anti-PD-1 APC (clone 29F-1A12), CD11b Alexa Fluor 700 (clone M1/70), anti-CD4 Brilliant Violet 570 (clone RM4-5), anti-CD44 PE-Cy7 (clone IM7), CD25 APC (clone 3C7) (Biolegend, San Diego, CA, USA), and CD11c FITC (clone HL3) (BD Biosciences, San José, CA, USA). The abs for intracellular staining were anti-FoxP3 Alexa Fluor 488 (clone MF23) (BD Biosciences) and anti-CTLA-4 PE (clone UC10-4F10-11) (Biolegend). A LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (Life Technologies, Thermo Scientific, Eugene, OR, USA) was used for dead cell exclusion. The abs used for intracellular cytokines evaluation were anti-IFNγ Alexa Fluor 700 (clone XMG1.2), TNFα PE-Cy7 (clone MP6-XT22), IL-2 FITC (clone JES6-5H4) (BD Biosciences), anti-perforin (clone S16009A), and anti-granzyme B (QA16A02) (Biolegend).
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