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567160Image1.png

Two-parameter 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 PE Mouse Anti-Human CD4 (Cat. No. 555347/561843/561844) and either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365; Left Panel) or Alexa Fluor® 647 Mouse Anti-IL-Human 22 antibody (Cat. No. 567160/567161; Right Panel) at 0.125 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.

567160-567161_fig2.png

Two-parameter 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 PE Mouse Anti-Human CD4 (Cat. No. 555347/561843/561844) and either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365; Left Panel) or Alexa Fluor® 647 Mouse Anti-IL-Human 22 antibody (Cat. No. 567160/567161; Right Panel) at 0.125 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.

567160Image1.png

Two-parameter 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 PE Mouse Anti-Human CD4 (Cat. No. 555347/561843/561844) and either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365; Left Panel) or Alexa Fluor® 647 Mouse Anti-IL-Human 22 antibody (Cat. No. 567160/567161; Right Panel) at 0.125 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.

567160Image1.png 567160-567161_fig2.png 567160Image1.png Red-Cap-Red-Label.jpg

Two-parameter 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 PE Mouse Anti-Human CD4 (Cat. No. 555347/561843/561844) and either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365; Left Panel) or Alexa Fluor® 647 Mouse Anti-IL-Human 22 antibody (Cat. No. 567160/567161; Right Panel) at 0.125 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.

Two-parameter 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 PE Mouse Anti-Human CD4 (Cat. No. 555347/561843/561844) and either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365; Left Panel) or Alexa Fluor® 647 Mouse Anti-IL-Human 22 antibody (Cat. No. 567160/567161; Right Panel) at 0.125 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.

Two-parameter 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 PE Mouse Anti-Human CD4 (Cat. No. 555347/561843/561844) and either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565365; Left Panel) or Alexa Fluor® 647 Mouse Anti-IL-Human 22 antibody (Cat. No. 567160/567161; Right Panel) at 0.125 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.

Product Details
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BD Pharmingen™
interleukin-22; IL-10-related T-cell-derived inducible factor; IL-TIF; ILTIF; IL-22a, ilTIFa; Iltif; TIFa
Human (QC Testing), Mouse (Tested in Development)
Mouse BALB/c IgG2a, κ
Human IL-22 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2916477
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

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Product Notices

  1. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  2. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  8. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  10. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  11. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
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567160 Rev. 1
Antibody Details
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MH22B2

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).

567160 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
Alexa Fluor™ 647
653 nm
669 nm
567160 Rev.1
Citations & References
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View product citations for antibody "567160" on CiteAb

Development References (8)

  1. 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
  2. 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
  3. 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
  4. Rutz S, Eidenschenk C, Ouyang W. IL-22, not simply a Th17 cytokine.. Immunol Rev. 2013; 252(1):116-32. (Biology). View Reference
  5. 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; 197(11):4473-4481. (Biology). View Reference
  6. 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
  7. 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
  8. Zenewicz LA. IL-22: There Is a Gap in Our Knowledge.. Immunohorizons. 2018; 2(6):198-207. (Biology). View Reference
View All (8) View Less
567160 Rev. 1

Please refer to Support Documents for Quality Certificates

 

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

 

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.