Cells were harvested and stained from the thymus and other organs. Data were acquired on LSR Fortessa or LSRII flow cytometers (BD Biosciences) and analyzed using software designed by the Division of Computer Research and Technology, NCI. Live cells were gated using forward scatter exclusion of dead cells stained with propidium iodide. The following antibodies were used for staining: TCRβ (H57–597), CD24 (30-F1), IL-7Rα (A7R34), NK1.1 (PK136), IL-2Rβ (TM-β1), IL-4Rα (M1), RORγt (Q31–378), T-bet (4B10), PLZF (9E12), and isotype control antibodies, all from eBioscience; CD44 (IM7), γc (4G3), CD4 (GK1.5 and RM4.5), and CD8α (53–6-7) from BD Biosciences; IL-21R (4A9) and CD45 (30-F11) from BioLegend. CD1d tetramers loaded with PBS-57 and unloaded controls were obtained from the NIH tetramer facility (Emory University, Atlanta, GA). Active caspase-3 was determined using the CaspGLOW™ fluorescein active caspase-3 staining kit (eBioscience).
Il 21r 4a9
Manufactured by BioLegend
IL-21R (4A9) is a mouse monoclonal antibody that recognizes the interleukin-21 receptor (IL-21R). IL-21R is a type I cytokine receptor that mediates the biological effects of interleukin-21 (IL-21).
Automatically generated - may contain errors
2 protocols using il 21r 4a9
1
Immunophenotyping of Thymic Subsets
2
Comprehensive Immune Cell Analysis
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Single cell suspensions were prepared from the thymus of indicated mice. Data were acquired using LSR Fortessa or LSRII flow cytometers (BD Biosciences) and analyzed using FlowJo. Live cells were gated by forward scatter exclusion of dead cells stained with propidium iodide. The following antibodies were used for staining: TCRβ (H57–597), HSA (30-F1), IL-7Rα (A7R34), NK1.1 (PK136), IL-2Rα (PC61.5), IL-2Rβ (TM-β1), IL-4Rα (M1), Foxp3 (FJK-16s), RORγt (AKFJS-9) and isotype control antibodies, all from eBioscience; TCRγδ (GL3), CD44 (IM7), γc (4G3), CD4 (GK1.5 and RM4.5), and CD8α (53-6-7) from BD Biosciences; IL-9Rα (RM9A4), Bcl-2 (BCL/10C4), PLZF (9E12), and IL-21R (4A9) from BioLegend. Fluorochome-conjugated CD1d tetramers loaded with PBS-567 and unloaded controls were obtained from the NIH tetramer facility (Emory University, Atlanta, GA). Intranuclear Foxp3, PLZF, and RORγt proteins were detected using a Foxp3 staining kit according to the manufacturer’s instructions (eBioscience). Active caspase-3 induction was determined using the CaspGLOW™ fluorescein active caspase-3 staining kit (eBioscience).
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