Human IL-12 Antibody

R&D Systems | Catalog # AF-219-NA

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Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Neutralization, Immunocytochemistry

Cited:

Immunohistochemistry, Western Blot, Neutralization, Immunocytochemistry, Affinity Chromatography, Bioassay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all IL-12 Primary Antibodies »

Product Specifications

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)

Specificity

Detects human IL-12 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human IL-12 Antibody

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

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.

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.
Human IL-12 / IL-23 p40 ELISA Standard Curve

Human IL-12 / IL-23 p40 ELISA Standard Curve

Recombinant Human IL‑12/IL‑23 p40 Monomer (Catalog # 309-IL) was serially diluted and captured by Mouse Anti-Human IL‑12/IL‑23 p40 Monoclonal Antibody (Catalog # MAB609) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human IL‑12 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-219-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)
Human IL-12 p70 ELISA Standard Curve

Human IL-12 p70 ELISA Standard Curve

Recombinant Human IL‑12 (Catalog # 219-IL) was serially diluted and captured by Mouse Anti-Human IL‑12 p70 Monoclonal Antibody (Catalog # MAB611R) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human IL‑12 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-219-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Applications for Human IL-12 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed human PBMCs

Western Blot

0.1 µg/mL
Sample: Recombinant Human IL‑12 (Catalog # 219-IL)

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.

Reviewed Applications

Read 1 review rated 5 using AF-219-NA in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

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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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

Alternate Names

CLMF1, IL12, IL12A, NKSF1

Entrez Gene IDs

3592 (Human); 16159 (Mouse); 84405 (Rat); 397053 (Porcine); 403977 (Canine); 102115316 (Cynomolgus Monkey); 493741 (Feline); 703205 (Rhesus Macaque)

Gene Symbol

IL12A

UniProt

P29459 (p35) & P29460 (p40)

Additional IL-12 Products

Product Documents for Human IL-12 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human IL-12 Antibody

For research use only

Citations for Human IL-12 Antibody

Customer Reviews for Human IL-12 Antibody (1)

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  • Human IL-12 Antibody
    Name: Michael Lehrke
    Application: Block/Neutralize
    Sample Tested: T cells
    Species: Human
    Verified Customer | Posted 08/10/2023
    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.
    Human IL-12 Antibody AF-219-NA

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