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      APC Rat Anti-Human IL-2
      554567Image1.png

      Expression of IL-2 by stimulated CD3+ and CD3-human PBMC. Human PBMC were stimulated for 6 h with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma, Cat. #C-9275), in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with FITC Mouse Anti-Human CD3 (Cat. No. 555332), fixed, permeabilized, and then stained with 0.25 µg of APC Rat Anti-Human IL-2 (Cat. No. 554567/561054; left panel).

      To demonstrate specificity of staining, the binding of APC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-2 (1.0 µg, Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with an excess of Purified Rat Anti-Human IL-2 (10 µg, Cat. No. 554563; right panel) prior to staining with the APC-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 unlabelled antibody blocking specificity controls (right panel). This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS™, FACS Vantage™ or FACSCalibur™.

      554567Image1.png

      Expression of IL-2 by stimulated CD3+ and CD3-human PBMC. Human PBMC were stimulated for 6 h with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma, Cat. #C-9275), in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with FITC Mouse Anti-Human CD3 (Cat. No. 555332), fixed, permeabilized, and then stained with 0.25 µg of APC Rat Anti-Human IL-2 (Cat. No. 554567/561054; left panel).

      To demonstrate specificity of staining, the binding of APC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-2 (1.0 µg, Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with an excess of Purified Rat Anti-Human IL-2 (10 µg, Cat. No. 554563; right panel) prior to staining with the APC-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 unlabelled antibody blocking specificity controls (right panel). This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS™, FACS Vantage™ or FACSCalibur™.

      18959A_554567_image3.png

      Expression of IL-2 by stimulated CD3+ and CD3-human PBMC. Human PBMC were stimulated for 6 h with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma, Cat. #C-9275), in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with FITC Mouse Anti-Human CD3 (Cat. No. 555332), fixed, permeabilized, and then stained with 0.25 µg of APC Rat Anti-Human IL-2 (Cat. No. 554567/561054; left panel).

      To demonstrate specificity of staining, the binding of APC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-2 (1.0 µg, Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with an excess of Purified Rat Anti-Human IL-2 (10 µg, Cat. No. 554563; right panel) prior to staining with the APC-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 unlabelled antibody blocking specificity controls (right panel). This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS™, FACS Vantage™ or FACSCalibur™.

      18959A_554567_image3.png

      Expression of IL-2 by stimulated CD3+ and CD3-human PBMC. Human PBMC were stimulated for 6 h with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma, Cat. #C-9275), in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with FITC Mouse Anti-Human CD3 (Cat. No. 555332), fixed, permeabilized, and then stained with 0.25 µg of APC Rat Anti-Human IL-2 (Cat. No. 554567/561054; left panel).

      To demonstrate specificity of staining, the binding of APC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-2 (1.0 µg, Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with an excess of Purified Rat Anti-Human IL-2 (10 µg, Cat. No. 554563; right panel) prior to staining with the APC-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 unlabelled antibody blocking specificity controls (right panel). This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS™, FACS Vantage™ or FACSCalibur™.

      554567Image1.png 554567Image1.png 18959A_554567_image3.png 18959A_554567_image3.png

      Expression of IL-2 by stimulated CD3+ and CD3-human PBMC. Human PBMC were stimulated for 6 h with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma, Cat. #C-9275), in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with FITC Mouse Anti-Human CD3 (Cat. No. 555332), fixed, permeabilized, and then stained with 0.25 µg of APC Rat Anti-Human IL-2 (Cat. No. 554567/561054; left panel).

      To demonstrate specificity of staining, the binding of APC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-2 (1.0 µg, Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with an excess of Purified Rat Anti-Human IL-2 (10 µg, Cat. No. 554563; right panel) prior to staining with the APC-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 unlabelled antibody blocking specificity controls (right panel). This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS™, FACS Vantage™ or FACSCalibur™.

      Expression of IL-2 by stimulated CD3+ and CD3-human PBMC. Human PBMC were stimulated for 6 h with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma, Cat. #C-9275), in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with FITC Mouse Anti-Human CD3 (Cat. No. 555332), fixed, permeabilized, and then stained with 0.25 µg of APC Rat Anti-Human IL-2 (Cat. No. 554567/561054; left panel).

      To demonstrate specificity of staining, the binding of APC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-2 (1.0 µg, Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with an excess of Purified Rat Anti-Human IL-2 (10 µg, Cat. No. 554563; right panel) prior to staining with the APC-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 unlabelled antibody blocking specificity controls (right panel). This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS™, FACS Vantage™ or FACSCalibur™.

      Expression of IL-2 by stimulated CD3+ and CD3-human PBMC. Human PBMC were stimulated for 6 h with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma, Cat. #C-9275), in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with FITC Mouse Anti-Human CD3 (Cat. No. 555332), fixed, permeabilized, and then stained with 0.25 µg of APC Rat Anti-Human IL-2 (Cat. No. 554567/561054; left panel).

      To demonstrate specificity of staining, the binding of APC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-2 (1.0 µg, Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with an excess of Purified Rat Anti-Human IL-2 (10 µg, Cat. No. 554563; right panel) prior to staining with the APC-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 unlabelled antibody blocking specificity controls (right panel). This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS™, FACS Vantage™ or FACSCalibur™.

      Expression of IL-2 by stimulated CD3+ and CD3-human PBMC. Human PBMC were stimulated for 6 h with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma, Cat. #C-9275), in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with FITC Mouse Anti-Human CD3 (Cat. No. 555332), fixed, permeabilized, and then stained with 0.25 µg of APC Rat Anti-Human IL-2 (Cat. No. 554567/561054; left panel).

      To demonstrate specificity of staining, the binding of APC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-2 (1.0 µg, Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with an excess of Purified Rat Anti-Human IL-2 (10 µg, Cat. No. 554563; right panel) prior to staining with the APC-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 unlabelled antibody blocking specificity controls (right panel). This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS™, FACS Vantage™ or FACSCalibur™.

      Product Details
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      BD Pharmingen™
      IL2; Interleukin-2; T-cell growth factor; TCGF
      Human (QC Testing)
      Rat IgG2a, κ
      Human IL-2 Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.2 mg/ml
      AB_398571
      Aqueous buffered solution containing ≤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 to APC under optimum conditions, and unconjugated antibody and free APC were removed.
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      Recommended Assay Procedures

      Immunofluorescent Staining and Flow Cytometric Analysis: The APC-conjugated MQ1-17H12 antibody (Cat. No. 554567) 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). For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.5 µg mAb/1X10^6 cells). For specific methodology, please visit the protocols section under "Cytokines (Intracellular Staining)" on our web

      site, http://www.bdbiosciences.com/us/s/resources.

      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; Cat No. 554603) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabelled 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 APC-R35-95 (Cat. No. 554690); use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/1X10^6 cells).

      Neutralization/Blocking: The NA/LE™ format of the MQ1-17H12 antibody (Cat. No. 554567) is useful for neutralization of human IL-2 bioactivity. A suitable NA/LE™ rat IgG2a isotype control to match the NA/LE™ MQ1-17H12 antibody is the R35-95 antibody, Cat. No. 554687.

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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
      6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      554567 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.

      554567 Rev. 4
      Format Details
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      APC
      Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 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
      APC
      651 nm
      660 nm
      554567 Rev.4
      Citations & References
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      View product citations for antibody "554567" 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
      554567 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.