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

(2 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 hybridoma culture supernatant
  • 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
    Allophycocyanin
Applications
  •  
    Recommended
    Concentration
    Sample
  • Intracellular Staining by Flow Cytometry
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 or Mouse Splenocytes by Flow Cytometry. (A) Human peripheral blood mononuclear cell (PBMCs) or (B) mouse splenocytes either untreated (light orange filled histogram) or treated with LPS (dark orange filled histogram) were stained with Mouse Anti-Human/Mouse IL‑12/IL‑35 p35 APC‑conjugated Monoclonal Antibody (Catalog # IC2191A) or isotype control antibody (Catalog # IC002A, 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
p35 is a 34-36 kDa monomeric member of the IL-6 superfamily of molecules. It is 219 amino acids in length, and possibly contains an N-terminal 22 aa signal sequence. In any event, it does not appear to be individually secreted, and its release from cells is commonly assumed to be as part of a disulfide-linked heterodimer linked to either p40 (forming IL-12) or EBI3 (forming IL-35). Multiple cell types are known to express p35, and include keratinocytes, CD138+ plasma cells, naïve B cells, monocyte-derived dendritic cells, macrophages, CD86lo B7-H1hi CD206+ B7-DC+ tolerogenic dendritic cells, neutrophils, activated monocytes and macrophages, trophoblasts, testicular endothelial cells, and T cells, including CD4+ and CD8+ T helper cells, and CD4+ CD25+ Tregs. The presence of the subunit does not necessarily indicate function, as the p35:p40 heterodimer is associated with promoting inflammatory IFN-gamma production by NK cells, while the p35:EBI3 heterodimer is anti-inflammatory through its inhibition of Th17 cell development. Over aa 23-219, human p35 shares 83% and 57% aa sequence identity with pig and mouse p35, respectively.
  • 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
Sample Type
  1. Differential regulation of interleukin-12 (IL-12)/IL-23 by Tim-3 drives T(H)17 cell development during hepatitis C virus infection.
    Authors: Wang, Jia M, Shi, Lei, Ma, Cheng J, Ji, Xiao J, Ying, Ruo S, Wu, Xiao Y, Wang, Ke S, Li, Guangyu, Moorman, Jonathan, Yao, Zhi Q
    J Virol, 2013;87(8):4372-83.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow Cytometry
  2. Regulatory T cells that recognize a ubiquitous stress-inducible self-antigen are long-lived suppressors of autoimmune arthritis.
    Authors: van Herwijnen M, Wieten L, van der Zee R, van Kooten P, Wagenaar-Hilbers J, Hoek A, den Braber I, Anderton S, Singh M, Meiring H, van Els C, van Eden W, Broere F
    Proc Natl Acad Sci U S A, 2012;109(35):14134-9.
    Species: Mouse
    Sample Type: Whole Cells
    Application: Flow Cytometry
Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG1 APC-conjugated Antibody

Ctrl IC002A 5
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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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