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Purified Hamster Anti-Mouse IL-12 Receptor β2
Purified Hamster Anti-Mouse IL-12 Receptor β2

Expression of cell surface IL-12R 2 by T helper cells. Mouse Th1 cell line, 2D6 (left panel) and Th2 cell line, D10 (center panel) were stained with purified anti-mouse IL-12 receptor ß2antibody (clone HAM10B9, 0.5 µg/test) followed by PE-conjugated anti-hamster IgG (0.25 µg, Cat . No. 554056). Staining with the HAM10B9 antibody (filled histograms) is compared to staining obtained using the isotype control antibody (open histograms). The histograms in the figure were derived from gated events with the forward and side light scatter characteristics of viable lymphocytes. Mouse splenocytes from C57BL/6 mice (right panel) were treated with an ammonium chloride lysing buffer to remove the red blood cells. Cells were subsequently cultured with ConA (2 µg/ml), PMA (50 ng/ml), Dextran sulfate (10 µg/ml), LPS (5 µg/ml), recombinant mouse IL-2 (10 ng/ml), recombinant mouse IL-12p70 (20 ng/ml) and anti-IL-4 antibody, clone 11B11 (5 µg/ml) for 5 days. Following culture the cells were harvested, washed, blocked with mouse Fc Block™ (Cat. No. 553141) and stained with purified anti-mouse IL-12 receptor β2 antibody (clone HAM10B9, 0.5 µg/test) followed by PE-conjugated anti-hamster IgG (0.25 µg, Cat . No. 554056) and Viaprobe (Cat. No. 555816). Staining with anti-mouse IL-12 receptor ß2antibody (clone HAM10B9, filled histograms) is compared to staining obtained using the isotype control antibody (Cat. No. 553969, open histograms). The histograms in the figure were derived from viable gated events (e.g. ViaProbe negative lymphocytes).

Expression of cell surface IL-12R 2 by T helper cells. Mouse Th1 cell line, 2D6 (left panel) and Th2 cell line, D10 (center panel) were stained with purified anti-mouse IL-12 receptor ß2antibody (clone HAM10B9, 0.5 µg/test) followed by PE-conjugated anti-hamster IgG (0.25 µg, Cat . No. 554056). Staining with the HAM10B9 antibody (filled histograms) is compared to staining obtained using the isotype control antibody (open histograms). The histograms in the figure were derived from gated events with the forward and side light scatter characteristics of viable lymphocytes. Mouse splenocytes from C57BL/6 mice (right panel) were treated with an ammonium chloride lysing buffer to remove the red blood cells. Cells were subsequently cultured with ConA (2 µg/ml), PMA (50 ng/ml), Dextran sulfate (10 µg/ml), LPS (5 µg/ml), recombinant mouse IL-2 (10 ng/ml), recombinant mouse IL-12p70 (20 ng/ml) and anti-IL-4 antibody, clone 11B11 (5 µg/ml) for 5 days. Following culture the cells were harvested, washed, blocked with mouse Fc Block™ (Cat. No. 553141) and stained with purified anti-mouse IL-12 receptor β2 antibody (clone HAM10B9, 0.5 µg/test) followed by PE-conjugated anti-hamster IgG (0.25 µg, Cat . No. 554056) and Viaprobe (Cat. No. 555816). Staining with anti-mouse IL-12 receptor ß2antibody (clone HAM10B9, filled histograms) is compared to staining obtained using the isotype control antibody (Cat. No. 553969, open histograms). The histograms in the figure were derived from viable gated events (e.g. ViaProbe negative lymphocytes).

Product Details
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BD Pharmingen™
Mouse (QC Testing)
Armenian Hamster IgG1, κ
Mouse IL-12Rβ2 transfectants
Flow cytometry (Routinely Tested)
0.5 mg/ml
AB_394475
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. Store undiluted at 4°C.

Recommended Assay Procedures

The purified anti-mouse IL-12 receptor ß2antibody (clone HAM10B9, Cat. No. 552819) can be used for the immunofluorescent staining (≤1 µg antibody/10e6 cells) and flow cytometric analysis of mouse Th1 or NK cells to measure their expressed levels of surface IL-12Rβ2. An appropriate purified immunoglobulin isotype control is clone A19-3 (Cat. No. 553969).

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
  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.
552819 Rev. 1
Antibody Details
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HAM10B9

The HAM10B9 monoclonal antibody specifically recognizes the β2subunit (IL-12Rβ2), of the mouse IL-12 receptor complex. The IL-12Rβ2 subunit associates with a β1 subunit to form a heterodimeric IL-12 receptor complex. Each one of the IL-12R subunits exhibits low affinity for IL-12, but in combination, they bind IL-12 with high affinity. The IL-12Rβ1 subunit interacts primarily with IL-12 p40 whereas the IL-12R β2 binds both to IL-12 p40 and IL-12 p35. IL-12Rβ1 is required for high affinity binding of IL-12 but IL-12R β2 is required for signaling.  The cytoplasmic regions of the β1 and β2 subunits contain the box 1 and box 2 motifs found in other cytokine receptors such as gp130, LIFR and G-CSFR. Naive T cells do not express IL-12R but both IL-12R subunits can be induced on T cells by antigenic stimulation. The IL12R is also expressed on activated NK cells. Th1 cells express both IL-12R subunits while Th2 cells lose the β2 subunit during differentiation. The HAM10B9 antibody was generated by immunizing hamsters with mouse IL-12Rβ2 transfectants.

552819 Rev. 1
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
552819 Rev.1
Citations & References
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Development References (10)

  1. Chakir H, Camilucci AA, Filion LG, Webb JR. Differentiation of murine NK cells into distinct subsets based on variable expression of the IL-12R beta 2 subunit. J Immunol. 2000 November; 165(9):4985-4993. (Biology). View Reference
  2. Gately MK, Renzetti LM, Magram J, et al. The interleukin-12/interleukin-12-receptor system: role in normal and pathologic immune responses. Annu Rev Immunol. 1998; 16:495-521. (Biology). View Reference
  3. Lúdvíksson BR, Ehrhardt RO, Strober W.. Role of IL-12 in intrathymic negative selection.. J Immunol. 1999 October; 163(8):4349-4359. (Clone-specific: Flow cytometry). View Reference
  4. Presky DH, Minetti LJ, Gillessen S, et al. Analysis of the multiple interactions between IL-12 and the high affinity IL-12 receptor complex. J Immunol. 1998; 160(5):2174-2179. (Biology). View Reference
  5. Presky DH, Yang H, Minetti LJ, et al. A functional interleukin 12 receptor complex is composed of two beta-type cytokine receptor subunits. Proc Natl Acad Sci U S A. 1996; 93(4):14002-14007. (Clone-specific). View Reference
  6. Smeltz RB, Chen J, Ehrhardt R, Shevach EM.. Role of IFN-gamma in Th1 differentiation: IFN-gamma regulates IL-18R alpha expression by preventing the negative effects of IL-4 and by inducing/maintaining IL-12 receptor beta 2 expression.. J Immunol. 2002 June; 168(12):6165-6172. (Clone-specific: Flow cytometry). View Reference
  7. Stahl N, Yancopoulos GD. The alphas, betas, and kinases of cytokine receptor complexes. Cell. 1993; 74(4):587-590. (Biology). View Reference
  8. Wang X, Wilkinson VL, Podlaski FJ, et al. Characterization of mouse interleukin-12 p40 homodimer binding to the interleukin-12 receptor subunits. Eur J Immunol. 1999; 29(6):2007-2013. (Biology). View Reference
  9. Wu C, Ferrante J, Gately MK, Magram J. Characterization of IL-12 receptor beta1 chain (IL-12Rbeta1)-deficient mice: IL-12Rbeta1 is an essential component of the functional mouse IL-12 receptor. J Immunol. 1997; 159(4):1658-1665. (Biology). View Reference
  10. Wu C, Wang X, Gadina M, O'Shea JJ, Presky DH, Magram J. IL-12 receptor beta 2 (IL-12R beta 2)-deficient mice are defective in IL-12-mediated signaling despite the presence of high affinity IL-12 binding sites. J Immunol. 2000; 165(11):6221-6228. (Biology). View Reference
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552819 Rev. 1

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