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Two color flow cytometric analysis of IL-2 expression by activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate (PMA, Sigma P-8139; 50 ng/ml) and Calcium Ionophore A23187 (Sigma C-9275; 1 μ g/ml), in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) [Cat. No. 554724]. The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656] and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) followed by washing and permeabilization using BD Perm/Wash™ Buffer (Cat. No. 554723). The permeabilized cells were then stained in BD Perm/Wash™ Buffer with PE Mouse Anti-Human CD3 antibody (Cat. No. 555333) and with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat No. 566928; Left Plot) or BD Horizon™ R718 Mouse Anti-Human IL-2 antibody (Cat No. 569490/569491; Right Plot). The pseudocolor density plot showing the correlated expression of IL-2 (or Ig Isotype control staining) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software.
BD Horizon™ R718 Mouse Anti-Human IL-2
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Companion Products
The 5344.111 antibody specifically binds to the multifunctional cytokine, human interleukin-2 (IL-2). IL-2 is produced by activated T cells and has multiple functions that can affect the growth, proliferation and differentiation of many different target cell types including T cells, B cells, NK cells, monocytes and macrophages. The immunogen used to generate the 5344.111 hybridoma was affinity-purified, recombinant human IL-2 protein.
Development References (7)
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Bitmansour AD, Douek DC, Maino VC, Picker LJ. Direct ex vivo analysis of human CD4(+) memory T cell activation requirements at the single clonotype level.. J Immunol. 2002; 169(3):1207-18. (Clone-specific: Flow cytometry, Intracellular Staining/Flow Cytometry). View Reference
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Jason J, Larned J. Single-cell cytokine profiles in normal humans: comparison of flow cytometric reagents and stimulation protocols. J Immunol Methods. 1997; 207(1):13-22. (Clone-specific: Flow cytometry, Intracellular Staining/Flow Cytometry). View Reference
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Liao W, Lin JX, Leonard WJ. IL-2 family cytokines: new insights into the complex roles of IL-2 as a broad regulator of T helper cell differentiation. Curr Protoc Immunol. 2011; 23(5):598-604. (Biology). View Reference
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Malek TR. The biology of interleukin-2. Annu Rev Immunol. 2008; 26:453-479. (Biology). View Reference
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Mendes R, Bromelow KV, Westby M. Flow cytometric visualisation of cytokine production by CD3-CD56+ NK cells and CD3+CD56+ NK-T cells in whole blood. Cytometry. 2000; 39(1):72-78. (Clone-specific: Flow cytometry, Intracellular Staining/Flow Cytometry). View Reference
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Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology: Intracellular Staining/Flow Cytometry). View Reference
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Waldmann TA. The biology of interleukin-2 and interleukin-15: implications for cancer therapy and vaccine design. Nat Rev Immunol. 2006; 6(8):595-601. (Biology). View Reference
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