Human/Mouse IL-12/IL-35 p35 PE-conjugated Antibody

(3 citations)   
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human IL‑12/IL-35 p35 in direct ELISAs and Western blots. Detects the p35 subunit either as part of a p40/p35 heterodimer or as a free subunit after reduction of the heterodimer. This antibody does not recognize IL-12 p40 homodimers but shows strong cross-reactivity with the p35 subunits from porcine and mouse systems.
  • Source
    Monoclonal Mouse IgG1 Clone # 27537
  • Purification
    Protein A or G purified from ascites
  • Immunogen
    S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL‑12/IL-35 p35
    Arg23-Ser219
    Accession # P29459
  • Formulation
    Supplied in a saline solution containing BSA and Sodium Azide.
  • Label
    Phycoerythrin
Applications
  •  
    Recommended
    Concentration
    Sample
  • Intracellular Staining by Flow Cytometry
    10 µL/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Intracellular Staining by Flow Cytometry

Detection of IL‑12/IL‑35 p35 in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) either untreated (orange open histogram) or treated with LPS (orange filled histogram) were stained with Mouse Anti-Human/Mouse IL‑12/IL‑35 p35 PE‑conjugated Monoclonal Antibody (Catalog # IC2191P) or isotype control antibody (Catalog # IC002P, blue open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Intracellular Staining by Flow Cytometry

Detection of IL‑12/IL‑35 p35 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes either untreated (orange open histogram) or treated with LPS (orange filled histogram) were stained with Mouse Anti-Human/Mouse IL‑12/IL‑35 p35 PE‑conjugated Monoclonal Antibody (Catalog # IC2191P) or isotype control antibody (Catalog # IC002P, blue open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Preparation and Storage
  • Shipping
    The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
  • Stability & Storage
    Protect from light. Do not freeze.
    • 12 months from date of receipt, 2 to 8 °C as supplied.
Background: IL-12/IL-35 p35
Interleukin 12 (IL-12) and interleukin 35 (IL-35) are heterodimeric cytokines composed of alpha and beta chains. IL-12 is composed of p35 and p40 subunits, while IL-35 is comprised of p35 paired with EBI-3 (1). In mice, IL-35 is produced by FoxP3+ regulatory T cells and may function as an inhibitory cytokine to suppress T cell proliferation (2). Human FoxP3+ Tregs do not constitutively express IL-35 (3), but expression may be induced by activated dendritic cells (4).
  • References:
    1. Collison, L.W. and D.A.A. Vignali (2008) Immunol. Rev. 226:248.
    2. Collison, L.W. et al. (2007) Nature 450:566.
    3. Bardel, E. et al. (2008) J. Immunol. 181:6898.
    4. Seyerl, M. et al.(2010) Eur. J. Immunol. 40:321.
  • Long Name:
    Interleukin 12 p35
  • Entrez Gene IDs:
    3592 (Human); 16159 (Mouse); 84405 (Rat)
  • Alternate Names:
    IL-12A
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

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Species
Applications
Sample Type
  1. IL-35 over-expression increases apoptosis sensitivity and suppresses cell growth in human cancer cells.
    Biochem Biophys Res Commun, 2013;430(1):364-9.
    Species: Human
    Sample Type: whole cells
    Application: Flow
  2. Generation of inducible immortalized dendritic cells with proper immune function in vitro and in vivo.
    Authors: Richter C, Thieme S, Bandola J, Laugsch M, Anastassiadis K, Brenner S
    PLoS ONE, 2013;8(4):e62621.
    Species: Mouse
    Sample Type: Whole Cells
    Application: Flow
  3. Cutting edge: Human regulatory T cells require IL-35 to mediate suppression and infectious tolerance.
    Authors: Chaturvedi V, Collison LW, Guy CS, Workman CJ, Vignali DA
    J. Immunol., 2011;186(12):6661-6.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow
Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG1 PE-conjugated Antibody

Ctrl IC002P 28
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Staining Reagents
Description Application Cat# Citations Images  

Flow Cytometry Permeabilization/Wash Buffer I (1X)

Flow FC005 4
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Flow Cytometry Fixation Buffer (1X)

Flow FC004 3
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Flow Cytometry Fixation & Permeabilization Buffer Kit I

Flow FC009
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Flow Cytometry Fixation/Permeabilization Buffer I (1X)

Flow FC007
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