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      PE Rat Anti-Human IL-2
      PE Rat Anti-Human IL-2
      Expression of IL-2 by stimulated CD3+ and CD3-human PBMC. Human PBMC were stimulated for 18 hours with PMA (Sigma, Cat. No. P-8139) and ionomycin (Sigma, Cat. No. I-0634), in the presence of BD GolgiStop™ (2 μl final concentration; Cat. No. 554724). The PBMC were stained with PE-Cy™5-anti-CD3 (Cat. No. 555334), fixed, permeabilized, and then stained with 20 μl of the PE Rat Anti-Human IL-2 antibody (Cat. No. 560902/559334/554566; left panel). To demonstrate specificity of staining, the binding of PE-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with recombinant human IL-2 (0.25 mg, Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with an excess of Purified Rat Anti-Human IL-2 (10 mg, Cat. No. 554563; right panel) prior to staining with the PE-MQ1-17H12 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking (middle panel) and unlabeled antibody blocking specificity controls (right panel).
      PE Rat Anti-Human IL-2
      Expression of IL-2 by stimulated CD3+ and CD3-human PBMC. Human PBMC were stimulated for 18 hours with PMA (Sigma, Cat. No. P-8139) and ionomycin (Sigma, Cat. No. I-0634), in the presence of BD GolgiStop™ (2 μl final concentration; Cat. No. 554724). The PBMC were stained with PE-Cy™5-anti-CD3 (Cat. No. 555334), fixed, permeabilized, and then stained with 20 μl of the PE Rat Anti-Human IL-2 antibody (Cat. No. 560902/559334/554566; left panel). To demonstrate specificity of staining, the binding of PE-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with recombinant human IL-2 (0.25 mg, Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with an excess of Purified Rat Anti-Human IL-2 (10 mg, Cat. No. 554563; right panel) prior to staining with the PE-MQ1-17H12 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking (middle panel) and unlabeled antibody blocking specificity controls (right panel).
      Product Details
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      BD Pharmingen™
      IL2; Interleukin-2; T-cell growth factor; TCGF
      Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
      Rat IgG2a, κ
      Human IL-2 Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      20 µl
      3558
      AB_397231
      Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
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      Recommended Assay Procedures

      Immunofluorescent Staining and Flow Cytometric Analysis: The PE-conjugated MQ1-17H12 antibody (Cat. No. 559334/560902/554566) can be used for multicolor immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-2-producing cells within mixed cell populations (see image). This 100 Test Size formulation of the PE-conjugated MQ1-17H12 antibody has been pre-titrated to assure effective intracellular detection of human IL-2 using 20 µl/1 × 10^6 cells.

      A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated MQ1-17H12 antibody with a molar excess of ligand (e.g., recombinant human IL-2; MQ1-17H12 antibody, Cat. No. 554563) prior to staining. A suitable rat IgG2a isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is also available in a 100 Test Size formulation PE-R35-95 (Cat. No. 559317). For specific methodology, please visit the protocols section on our web site, http://www.bdbiosciences.com/resources/index.jsp

      Important Note: This pretitered antibody solution does not contain a cell permeabilization agent. It is necessary to include a cell permeabilization agent when using the pre-titered antibody solution to stain fixed and permeabilized cells. BD Perm/Wash™ Buffer (Cat. No 554723) contains the

      permeabilization agent saponin and is useful for this purpose as described in the USAGE section on the following page.

      Usage

      1. Resuspend 1 × 10^6 fixed and permeabilized cells in 20 µl of the pre-titered antibody solution and 30 µl of 1× BD Perm/Wash™ Buffer (Cat. No. 554723).

      2. Incubate the cell suspension for 15 minutes (at RT or 4°C).

      3. Wash twice in 100 µl of 1× BD Perm/Wash™ Buffer (Cat. No. 554723).

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      Product Notices

      1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
      2. An isotype control should be used at the same concentration as the antibody of interest.
      3. 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.
      4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      7. Cy is a trademark of GE Healthcare.
      8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      559334 Rev. 4
      Antibody Details
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      MQ1-17H12

      The MQ1-17H12 monoclonal 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, differentiation and survival of many different target cell types including T cells, B cells, NK cells, monocytes and macrophages. The immunogen used to generate the MQ1-17H12 hybridoma was purified recombinant human IL-2 protein. The MQ1-17H12 antibody reportedly neutralizes the biological activity of human IL-2.

      559334 Rev. 4
      Format Details
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      PE
      R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      PE
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      576 nm
      559334 Rev.4
      Citations & References
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      View product citations for antibody "559334" on CiteAb

      Development References (2)

      1. Abrams J. Immunoenzymetric assay of mouse and human cytokines using NIP-labeled anti-cytokine antibodies. Curr Protoc Immunol. 2001; 1:6.20-6.21. (Biology). View Reference
      2. Abrams JS, Roncarolo MG, Yssel H, Andersson U, Gleich GJ, Silver JE. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 1992; 127:5-24. (Biology). View Reference
      559334 Rev. 4

       

      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.