Human IL-12 Biotinylated Antibody

(8 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human IL-12 in ELISAs and Western blots. When used in combination with the monoclonal capture antibody anti‑human IL-12 p40 (Catalog # MAB609) in sandwich ELISAs, less than 2.5% cross‑reactivity was observed with rhIL-12 p70. When used in combination with the monoclonal capture antibody anti-human IL-12 p70 (Catalog # MAB611) in sandwich ELISAs, less than 0.1% cross‑reactivity was observed with rhIL‑12 p40, rhIL‑12 p35 and rmIL‑12.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL‑12 heterodimer
    Arg23-Ser219 of p35; Ile23-Ser328 of p40
    Accession # P29459 (p35) & P29460 (p40)
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
  • Label
    Biotin
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Human IL‑12 (Catalog # 219-IL)
  • Immunocytochemistry
    5-15 µg/mL
    See below
    • Human IL-12 p70 Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    2-8 µg/mL 
    Human IL‑12 p70 Antibody (Catalog # MAB611)
  • ELISA Capture (Matched Antibody Pair)
    2-8 µg/mL 
    Human IL‑12 p70 Antibody (Catalog # MAB611R)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Human IL‑12 Biotinylated Antibody (Catalog # BAF219)
  • ELISA Standard
     
    Recombinant Human IL-12 Protein (Catalog # 219-IL)
    • Human IL-12/IL-23 p40 Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    2-8 µg/mL 
    Human IL‑12/IL‑23 p40 Antibody (Catalog # MAB609)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Human IL‑12 Biotinylated Antibody (Catalog # BAF219)
  • ELISA Standard
     
    Recombinant Human IL-12/IL-23 p40 Monomer Protein (Catalog # 309-IL)
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
Immunocytochemistry
IL‑12 in Human PBMCs. IL‑12 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with calcium ionomycin and PMA using Goat Anti-Human IL‑12 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF219) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; Catalog # NL999) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: IL-12

Interleukin 12, also known as natural killer cell stimulatory factor (NKSF) or cytotoxic lymphocyte maturation factor (CLMF), is a pleiotropic cytokine originally identified in the medium of activated human B lymphoblastoid cell lines. The p40 subunit of IL-12 has been shown to have extensive amino acid sequence homology to the extracellular domain of the human IL-6 receptor while the p35 subunit shows distant but significant sequence similarity to IL-6, G-CSF, and chicken MGF. These observations have led to the suggestion that IL-12 might have evolved from a cytokine/soluble receptor complex. Human and murine IL-12 share 70% and 60% amino acid sequence homology in their p40 and p35 subunits, respectively. IL-12 apparently shows species specificity with human IL-12 reportedly showing minimal activity in the murine system.

IL-12 is produced by macrophages and B lymphocytes and has been shown to have multiple effects on T cells and natural killer (NK) cells. These effects include inducing production of IFN-gamma and TNF by resting and activated T and NK cells, synergizing with other IFN-gamma  inducers at both the transcriptional and post-transcriptional levels. This interaction induces IFN-gamma  gene expression, enhancing the cytotoxic activity of resting NK and T cells, inducing and synergizing with IL-2 in the generation of lymphokine-activated killer (LAK) cells, acting as a co-mitogen to stimulate proliferation of resting T cells, and inducing proliferation of activated T and NK cells. Evidence indicates that IL-12, produced by macrophages in response to infectious agents, is a central mediator of the cell-mediated immune response by its actions on the development, proliferation, and activities of TH1 cells. In its role as the initiator of cell-mediated immunity, it has been suggested that IL-12 has therapeutic potential as a stimulator of cell-mediated immune responses to microbial pathogens, metastatic cancers, and viral infections such as AIDS.

  • Long Name:
    Interleukin 12
  • Entrez Gene IDs:
    3592 (Human); 16159 (Mouse); 84405 (Rat); 403977 (Canine); 493741 (Feline)
  • Alternate Names:
    CLMF p35; CLMF1; Cytotoxic lymphocyte maturation factor 35 kDa subunit; IL-12 subunit p35; IL12; IL-12; IL-12, subunit p35; IL12A; interleukin 12, p35; interleukin 12A (natural killer cell stimulatory factor 1, cytotoxic lymphocytematuration factor 1, p35); interleukin-12 alpha chain; interleukin-12 subunit alpha; natural killer cell stimulatory factor 1, 35 kD subunit; NF cell stimulatory factor chain 1; NK cell stimulatory factor chain 1; NKSF1; p35
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.

8 Citations: Showing 1 - 8
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Species
Applications
Sample Type
  1. TLR-stimulated CD34 stem cell-derived human skin-like and monocyte-derived dendritic cells fail to induce Th17 polarization of naive T cells but do stimulate Th1 and Th17 memory responses.
    Authors: Duraisingham SS, Hornig J, Gotch F, Patterson S
    J. Immunol., 2009;183(4):2242-51.
    Species: Human
    Sample Type: Whole Cells
    Application: ELISA Development
  2. Upregulation of human cytomegalovirus by HIV type 1 in human lymphoid tissue ex vivo.
    Authors: Biancotto A, Iglehart SJ, Lisco A, Vanpouille C, Grivel JC, Lurain NS, Reichelderfer PS, Margolis LB
    AIDS Res. Hum. Retroviruses, 2008;24(3):453-62.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Luminex Development
  3. Abnormal activation and cytokine spectra in lymph nodes of people chronically infected with HIV-1.
    Authors: Biancotto A, Grivel JC, Iglehart SJ, Vanpouille C, Lisco A, Sieg SF, Debernardo R, Garate K, Rodriguez B, Margolis LB, Lederman MM
    Blood, 2007;109(10):4272-9.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Luminex Development
  4. HIV-1 pathogenesis differs in rectosigmoid and tonsillar tissues infected ex vivo with CCR5- and CXCR4-tropic HIV-1.
    Authors: Grivel JC, Elliott J, Lisco A, Biancotto A, Condack C, Shattock RJ, McGowan I, Margolis L, Anton P
    AIDS, 2007;21(10):1263-72.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Luminex Development
  5. Effect of serum content and diluent selection on assay sensitivity and signal intensity in multiplex bead-based immunoassays.
    Authors: Pfleger C, Schloot N, ter Veld F
    J. Immunol. Methods, 2007;329(1):214-8.
    Species: Human
    Sample Type: Serum
    Application: Luminex Development
  6. Blockade of TLR9 agonist-induced type I interferons promotes inflammatory cytokine IFN-gamma and IL-17 secretion by activated human PBMC.
    Authors: Meyers JA, Mangini AJ, Nagai T, Roff CF, Sehy D, van Seventer JM
    Cytokine, 2006;35(5):235-46.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Development
  7. Human tumor membrane vesicles modified to express glycolipid-anchored IL-12 by protein transfer induce T cell proliferation in vitro: a potential approach for local delivery of cytokines during vaccination.
    Authors: Nagarajan S, Selvaraj P
    Vaccine, 2005;24(13):2264-74.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  8. Direct comparison of traditional ELISAs and membrane protein arrays for detection and quantification of human cytokines.
    Authors: Copeland S, Siddiqui J, Remick D
    J. Immunol. Methods, 2004;284(1):99-106.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Array Development
Isotype Controls
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Normal Goat IgG Biotinylated Control

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EvenCoat Streptavidin Coated Plates, 5 Pack

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