PE Mouse Anti-Mouse CD212

BD Pharmingen™ PE Mouse Anti-Mouse CD212

Name: PE Mouse Anti-Mouse CD212
Brand: BD Pharmingen™
SKU: 551974

Clone 114

(RUO)

Expression of cell surface IL-12Rβ1 by LAK cells. Mouse splenocytes from C57BL/6 mice were treated with an ammonium chloride lysing buffer to remove the red blood cells. Cells were subsequently cultured with 3000 U/ml of mouse IL-2 for 3-4 days at 37°C. At 3-4 days the adherent fraction of the cells was separated from the non-adherent fraction and fresh IL-2 was added (3000 U/ml) for an additional 3-4 days. Following culture both of the adherent and non-adherent cells were harvested, washed, blocked with mouse Fc Block™ (Cat. No. 553141) and stained with R-PE-conjguated 114 antibody (1 µg/test, Cat. No. 551974). Staining with the 114 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 light scattering characteristics of viable lymphocytes.

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Product Details

Brand: BD Pharmingen™

Alternative Name: Il12rb1; IL-12R-beta-1; IL-12 Receptor β1 chain

Reactivity: Mouse (QC Testing)

Isotype: Mouse IgG2a, κ

Immunogen: IL-12Rβ1 Transfected Cell Line

Application: Flow cytometry/immunoassay (Routinely Tested)

Concentration: 0.2 mg/ml

Entrez Gene ID: 16161

RRID: AB_394310

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

Recommended Assay Procedure:

Immunofluorescent Staining and Flow Cytometric Analysis. The R-PE-conjugated 114 antibody (Cat. No. 551974) can be used for the immunofluorescent staining (≤ 1 µg antibody/10e6 cells) and flow cytometric analysis of muose T cells and NK cells to measure their expressed levels of surface IL-12Rβ1. An appropriate PE-conjugated immunoglobulin isotype control is clone G155-178 (Cat. No. 553457).

Product Notices

Since applications vary, each investigator should titrate the reagent to obtain optimal results.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.

551974 Rev. 1

Antibody Details

114

The 114 monoclonal antibody specifically binds to mouse CD212 (the β1 subunit of IL-12Rβ1), originally termed IL-12Rβ, of the mouse IL-12 receptor complex. The IL-12Rβ1 subunit associates with a β2 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-12Rβ2 binds both to IL-12 p40 and IL-12 p35. IL-12Rβ1 is required for high affinity binding of IL-12 but IL-12Rβ2 is required for signaling. IL-12Rβ1 has more recently been described to bind IL-23, a heterodimer formed of the p40 subunit from IL-12, and p19. The cytoplasmic regions of the β1 and β2 subunits contain the box1 and box2 motifs found in other cytokine receptors such as gp130, LIFR and G-CSFR. IL-12Rβ1 are primarily expressed by activated T cells and NK cells. Experiments with IL-12Rβ1 deficient mice have shown that IL-12Rβ1 is necessary for mouse T and NK cell responsiveness to IL-12 p75. The 114 antibody was generated by immunizing IL-12Rβ1 deficient mice of (129 x BALB/c)F1 background with mouse Ba/F3 cells that were stably transfected with IL-12Rβ1.

This antibody is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.

551974 Rev. 1

Format Details

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.

Format: PE

Excitation Source: Yellow-Green 488 nm, 532 nm, 561 nm

Excitation Max: 496 nm, 566 nm

Emission Max: 576 nm

551974 Rev.1

Citations & References

View product citations for antibody "551974" on CiteAb

Development References (8)

Chua AO, Wilkinson VL, Presky DH, Gubler U. Cloning and characterization of a mouse IL-12 receptor-beta component. J Immunol. 1995; 155(9):4286-4294. (Clone-specific). View Reference
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
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
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
Stahl N, Yancopoulos GD. The alphas, betas, and kinases of cytokine receptor complexes. Cell. 1993; 74(4):587-590. (Biology). View Reference
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
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
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

View All (8)

551974 Rev. 1

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