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Human IL-12 Antibody

Catalog #: AF-219-NA
19 Citations 1 Review Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF-219-NA
AF-219-SP
Cell Proliferation Induced by IL‑12 and Neutralization by Human IL‑12 Antibody.
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Citations (19)
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Reviews (1)
Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human IL-12 Antibody Summary

Species Reactivity
Human
Specificity
Detects human IL-12 in direct ELISAs and Western blots.
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 Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Human IL‑12 (Catalog # 219-IL)
Immunocytochemistry
5-15 µg/mL
Immersion fixed human PBMCs
Neutralization
Measured by its ability to neutralize IL‑12-induced proliferation in PHA-activated human peripheral blood mononuclear cells (PBMC). Yokota, T. et al. (1986) Proc. Natl. Acad. Sci. USA 83:5894. The Neutralization Dose (ND50) is typically 5-50 ng/mL in the presence of 1 ng/mL Recombinant Human IL‑12.

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.

Scientific Data

Neutralization Cell Proliferation Induced by IL‑12 and Neutralization by Human IL‑12 Antibody. View Larger

Cell Proliferation Induced by IL‑12 and Neutralization by Human IL‑12 Antibody. Recombinant Human IL-12 (219-IL) stimulates proliferation in PHA-activated human peripheral blood mononuclear cells (PBMC) in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human IL-12 (1 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL-12 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-219-NA). The ND50 is typically 5-50 ng/mL.

Immunocytochemistry View Larger

Detection of IL‑12 in Human PBMCs. IL‑12 was detected in immersion fixed Human PBMCs using Goat Anti-Human IL‑12 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-219-NA) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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. Current 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); 397053 (Porcine); 403977 (Canine); 102115316 (Cynomolgus Monkey); 493741 (Feline); 703205 (Rhesus Macaque)
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

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Citations for Human IL-12 Antibody

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.

19 Citations: Showing 1 - 10
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  1. A novel microRNA-182/Interleukin-8 regulatory axis controls osteolytic bone metastasis of lung cancer
    Authors: Ming-Na Zhao, Ling-Fei Zhang, Zhen Sun, Li-Hua Qiao, Tao Yang, Yi-Zhe Ren et al.
    Cell Death & Disease
  2. The p40 subunit of interleukin (IL)-12 promotes stabilization and export of the p35 subunit: implications for improved IL-12 cytokine production
    Authors: Rashmi Jalah, Margherita Rosati, Brunda Ganneru, Guy R. Pilkington, Antonio Valentin, Viraj Kulkarni et al.
    Journal of Biological Chemistry
  3. Fungal sensing by dectin-1 directs the non-pathogenic polarization of TH17 cells through balanced type I IFN responses in human DCs
    Authors: SI Gringhuis, TM Kaptein, EBM Remmerswaa, A Drewniak, BA Wevers, B Theelen, GRAM D'Haens, T Boekhout, TBH Geijtenbee
    Nature Immunology, 2022-12-01;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  4. Inhibition of G-protein betagamma signaling enhances T cell receptor-stimulated interleukin 2 transcription in CD4+ T helper cells.
    Authors: Yost E, Hynes T, Hartle C, Ott B, Berlot C
    PLoS ONE, 2015-01-28;10(1):e0116575.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  5. Mycobacterium bovis Bacillus Calmette-Guerin suppresses inflammatory Th2 responses by inducing functional alteration of TSLP-activated dendritic cells.
    Authors: Yokoi T, Amakawa R, Tanijiri T, Sugimoto H, Torii Y, Amuro H, Son Y, Tajima K, Liu YJ, Ito T, Fukuhara S
    Int. Immunol., 2008-08-13;20(10):1321-9.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  6. Upregulated IL-23 and IL-17 in Behcet patients with active uveitis.
    Authors: Chi W, Zhu X, Yang P, Liu X, Lin X, Zhou H, Huang X, Kijlstra A
    Invest. Ophthalmol. Vis. Sci., 2008-07-01;49(7):3058-64.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  7. Distinct subsets of human invariant NKT cells differentially regulate T helper responses via dendritic cells.
    Authors: Liu TY, Uemura Y, Suzuki M, Narita Y, Hirata S, Ohyama H, Ishihara O, Matsushita S
    Eur. J. Immunol., 2008-04-01;38(4):1012-23.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  8. Statin-induced proinflammatory response in mitogen-activated peripheral blood mononuclear cells through the activation of caspase-1 and IL-18 secretion in monocytes.
    Authors: Coward WR, Marei A, Yang A, Vasa-Nicotera MM, Chow SC
    J. Immunol., 2006-05-01;176(9):5284-92.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  9. Innate immune responses to herpes simplex virus type 2 influence skin homing molecule expression by memory CD4+ lymphocytes.
    Authors: Koelle DM, Huang J, Hensel MT, McClurkan CL
    J. Virol., 2006-03-01;80(6):2863-72.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  10. 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, 2005-12-07;24(13):2264-74.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Affinity Chromatography
  11. Effect of piceatannol in human monocyte-derived dendritic cells in vitro.
    Authors: Takei M, Umeyama A, Arihara S, Matsumoto H
    J Pharm Sci, 2005-05-01;94(5):974-82.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  12. Dendritic cells induce MUC1 expression and polarization on human T cells by an IL-7-dependent mechanism.
    Authors: Vasir B, Avigan D, Wu Z, Crawford K, Turnquist S, Ren J, Kufe D
    J. Immunol., 2005-02-15;174(4):2376-86.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  13. Natural and induced CD4+CD25+ cells educate CD4+CD25- cells to develop suppressive activity: the role of IL-2, TGF-beta, and IL-10.
    Authors: Zheng SG, Wang JH, Gray JD, Soucier H, Horwitz DA
    J. Immunol., 2004-05-01;172(9):5213-21.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  14. NK cells regulate CD8+ T cell effector function in response to an intracellular pathogen.
    Authors: Vankayalapati R, Klucar P, Wizel B, Weis SE, Samten B, Safi H, Shams H, Barnes PF
    J. Immunol., 2004-01-01;172(1):130-7.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  15. APC-independent activation of NK cells by the Toll-like receptor 3 agonist double-stranded RNA.
    Authors: Schmidt KN, Leung B, Kwong M, Zarember KA, Satyal S, Navas TA, Wang F, Godowski PJ
    J. Immunol., 2004-01-01;172(1):138-43.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  16. Activation of a subset of human NK cells upon contact with Plasmodium falciparum-infected erythrocytes.
    Authors: Artavanis-Tsakonas K, Eleme K, McQueen KL, Cheng NW, Parham P, Davis DM, Riley EM
    J. Immunol., 2003-11-15;171(10):5396-405.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  17. Cytokine profile of the rheumatoid nodule suggests that it is a Th1 granuloma.
    Authors: Hessian PA, Highton J, Kean A, Sun CK, Chin M
    Arthritis Rheum., 2003-02-01;48(2):334-8.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  18. Evidence for expression of eosinophil-associated IL-12 messenger RNA and immunoreactivity in bronchial asthma.
    Authors: Nutku E, Gounni AS, Olivenstein R, Hamid Q
    J. Allergy Clin. Immunol., 2000-08-01;106(2):288-92.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  19. TSLP-activated dendritic cells induce an inflammatory T helper type 2 cell response through OX40 ligand
    Authors: Tomoki Ito, Yui-Hsi Wang, Omar Duramad, Toshiyuki Hori, Guy J. Delespesse, Norihiko Watanabe et al.
    The Journal of Experimental Medicine

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Human IL-12 Antibody
By Michael Lehrke on 08/10/2023
Application: B/N Sample Tested: T cells Species: Human

Utilized during in vitro Th2 polarization cultures to prevent unwanted differentiation of IFNg-producing Th1 cells.

Naive CD4 T cells were stimulated with anti-CD3/anti-CD28 antibodies and polarized with culture media that included 20 ng/mL IL-4, 5 ug/mL anti-IL-12 antibody, and 5 ug/mL anti-IFNg antibody. After 5 days of culture, supernatants were analyzed AlphaLISA to assess IL-5 and IFNg production.

As expected, the Th2 cultures did not produce unwanted IFNg indicating Th1 polarization was inhibited.

Block/Neutralize Human IL-12 Antibody AF-219-NA