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Multicolor flow cytometric analysis of IL-22 expression in human peripheral blood mononuclear cells (PBMCs). PBMCs were cultured in complete tissue culture medium with Phorbol 12-Myristate 13-Acetate (PMA; Sigma, 50 ng/ml) and Ionomycin (Sigma, 500 ng/ml) in the presence of BD GolgiPlug™ Protein Transport Inhibitor (Cat. No. 555029) for 6 hours. The cells were harvested, then fixed and permeabilized using BD Cytofix™ Fixation Buffer (Cat. No. 554655) and BD Perm/Wash™ buffer (Cat. No. 554723). The cells were then stained with BD Horizon™ BUV395 Mouse Anti-Human CD4 (Cat. No. 564724) and either BD Horizon™ R718 Mouse IgG2a, κ Isotype Control (Cat. No. 567089; Left Plot) or BD Horizon™ R718 Mouse anti-Human IL-22 antibody (Cat. No. 567569/567568; Right Plot) at 0.015 µg/test. Bivariate pseudocolor density plots showing the correlated expression of IL-22 (or Ig Isotype control staining) versus CD4 were 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 X-20 LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ R718 Mouse Anti-Human IL-22
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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
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The MH22B2 monoclonal antibody specifically recognizes human Interleukin-22 (IL-22), which is encoded by the IL22 gene. Cross-reactivity of MH22B2 mAb to mouse IL-22 (which shares 79% amino acid identity with human IL-22) has been observed by ELISA and by flow cytometry of HEK293 cells transfected with mouse IL-22 and TH17-differentiated CD4-positive T cells from C57BL/6 mice. IL-22 (with or without other cytokines) is secreted by many T-cell and innate-lymphoid-cell populations. Evidence that IL-22 plays a key role in mucosal immunity includes the restricted expression of the alpha subunit of the heterodimeric IL-22 receptor, called IL-22R1, on epithelial cells and cells of epithelial origin. At epithelial surfaces, IL-22 elicits antimicrobial defenses and maintains epithelial integrity. Alternatively, uncontrolled IL-22 production can result in certain inflammatory disorders. Regulation of IL-22 expression is complex, involving other cytokines (eg, IL-6, IL-23, and TGF-β) and many transcription factors, (eg, AHR, c-Maf, STAT3, RORɤT, BATF, and others).
The antibody was conjugated to BD Horizon™ Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon™ Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.
Development References (8)
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Colonna M. Interleukin-22-producing natural killer cells and lymphoid tissue inducer-like cells in mucosal immunity.. Immunity. 2009; 31(1):15-23. (Biology). View Reference
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Le-Thi-Phuong T, Dumoutier L, Renauld JC, Van Snick J, Coutelier JP. Divergent roles of IFNs in the sensitization to endotoxin shock by lactate dehydrogenase-elevating virus.. Int Immunol. 2007; 19(11):1303-11. (Clone-specific: ELISA). View Reference
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Martin B, Hirota K, Cua DJ, Stockinger B, Veldhoen M. Interleukin-17-producing gammadelta T cells selectively expand in response to pathogen products and environmental signals.. Immunity. 2009; 31(2):321-30. (Clone-specific: Flow cytometry). View Reference
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Rutz S, Eidenschenk C, Ouyang W. IL-22, not simply a Th17 cytokine.. Immunol Rev. 2013; 252(1):116-32. (Biology). View Reference
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Suurmond J, Habets KL, Dorjée AL, Huizinga TW, Toes RE. Expansion of Th17 Cells by Human Mast Cells Is Driven by Inflammasome-Independent IL-1β. J Immunol. 2016; 164(11):4473-4481. (Biology). View Reference
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Trifari S, Kaplan CD, Tran EH, Crellin NK, Spits H. Identification of a human helper T cell population that has abundant production of interleukin 22 and is distinct from T(H)-17, T(H)1 and T(H)2 cells.. Nat Immunol. 2009; 10(8):864-71. (Biology). View Reference
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Veldhoen M, Hirota K, Westendorf AM, et al. The aryl hydrocarbon receptor links TH17-cell-mediated autoimmunity to environmental toxins.. Nature. 2008; 453(7191):106-9. (Immunogen: Flow cytometry). View Reference
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Zenewicz LA. IL-22: There Is a Gap in Our Knowledge.. Immunohorizons. 2018; 2(6):198-207. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.