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Purified Rat Anti-Human IL-2
Purified Rat Anti-Human IL-2
Expression of IL-2 by stimulated CD3+ human PBMC. Human PBMCs were stimulated for 6 hours with PMA (50 ng/ml final concentration; Sigma, Cat. #P-8139) and calcium ionophore A23187 (500-100 ng/ml final concentration; Sigma, Cat. #C-9275) in the presence of BD GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMCs were stained with PE-Cy™5-anti-CD3 (Cat. No. 555334), fixed, permeabilized, and then stained with 0.25 µg of FITC-rat anti-human IL-2 antibody (Cat. No. 554565) using Pharmingen's staining protocol (left panel). To demonstrate specificity of staining, the binding of FITC-MQ1-17H12 was blocked by the preincubation of the fluorochrome - conjugated antibody with recombinant human IL-2 (10 ng/ml final concentration; Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with Purified Rat Anti-Human IL-2 (5.0 µg, Cat. No. 554563; right panel) prior to staining with the FITC-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 (right panel) specificity controls.
Expression of IL-2 by stimulated CD3+ human PBMC. Human PBMCs were stimulated for 6 hours with PMA (50 ng/ml final concentration; Sigma, Cat. #P-8139) and calcium ionophore A23187 (500-100 ng/ml final concentration; Sigma, Cat. #C-9275) in the presence of BD GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMCs were stained with PE-Cy™5-anti-CD3 (Cat. No. 555334), fixed, permeabilized, and then stained with 0.25 µg of FITC-rat anti-human IL-2 antibody (Cat. No. 554565) using Pharmingen's staining protocol (left panel). To demonstrate specificity of staining, the binding of FITC-MQ1-17H12 was blocked by the preincubation of the fluorochrome - conjugated antibody with recombinant human IL-2 (10 ng/ml final concentration; Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with Purified Rat Anti-Human IL-2 (5.0 µg, Cat. No. 554563; right panel) prior to staining with the FITC-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 (right panel) specificity controls.
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
ELISA Capture (Routinely Tested), Intracellular block/flow cytometry (Tested During Development)
0.5 mg/ml
3558
AB_398570
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Blocking Control for Intracellular Staining: The purified MQ1-17H12 antibody (Cat. No. 554563) can be used as a blocking control to demonstrate specificity of IL-2 staining of directly conjugated MQ1-17H12 antibody. To perform this control, the fixed/permeabilized cells (~ 1 million) can be incubated with 1 -10 µg of unlabeled MQ1-17H12 antibody (Cat. No. 554563) for 20 minutes at 4°C, prior to staining with conjugated antibody (e.g., 0.1 -0.5 µg mAb/1X10^6 cells). The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. For specific methodology, please visit the protocol section on our web site, http://www.bdbiosciences.com/resources/index.jsp

Neutralization/Blocking: THE NA/LE™ format of the MQ1-17H12 antibody (Cat. 554562) is useful for neutralization of human IL-2 bioactivity. A suitable NA/LE™ isotype control to match the NA/LE™ MQ1-17H12 antibody is the R35-95 antibody (Cat. No. 554687).

ELISA Capture: The purified MQ1-17H12 antibody is useful as a capture antibody for a sandwich ELISA for specifically measuring human

IL-2 protein levels. The biotinylated B33-2 antibody (Cat. No. 555040) can be paired with the purified MQ1-17H12 antibody (Cat. No. 554563) as the capture antibody, with recombinant human IL-2 (Cat. No. 554603) as the standard. For specific methodology, please visit the protocols section on our website, http://www.bdbiosciences.com/resources/index.jsp

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. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. Cy is a trademark of GE Healthcare.
  6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
554563 Rev. 6
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.

554563 Rev. 6
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
554563 Rev.6
Citations & References
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Development References (3)

  1. Andersson J, Abrams J, Bjork L, et al. Concomitant in vivo production of 19 different cytokines in human tonsils. Immunology. 1994; 83(1):16-24. (Biology). View Reference
  2. Fernandez V, Andersson J, Andersson U, Troye-Blomberg M. Cytokine synthesis analyzed at the single-cell level before and after revaccination with tetanus toxoid. Eur J Immunol. 1994; 24(8):1808-1815. (Biology). View Reference
  3. 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: IC/FCM Block). View Reference
554563 Rev. 6

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