Interleukin 12, also known as natural killer cell stimulatory factor (NKSF) or cytotoxic lymphocyte maturation factor (CLMF), is a pleiotrophic 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.
Human IL-12 p70 Antibody (24945R)
R&D Systems | Catalog # MAB611R
Recombinant Monoclonal Antibody.
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
ELISA Capture (Matched Antibody Pair), Immunocytochemistry
Cited:
Bioassay
Label
Unconjugated
Antibody Source
Recombinant Monoclonal Mouse IgG1 Clone # 24945R
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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)
Arg23-Ser219 of p35, Ile23-Ser328 of p40
Accession # P29459(p35) & P29460 (p40)
Specificity
Detects human IL-12 p70 in ELISAs. In sandwich immunoassays, no cross-reactivity with recombinant human (rh) IL-12 p40, rhIL-12 p35, rhIL-23 or recombinant mouse IL-12 is observed.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG1
Scientific Data Images for Human IL-12 p70 Antibody (24945R)
IL‑12 p70 in THP‑1 Human Cell Line.
IL‑12 p70 was detected in immersion fixed THP‑1 human acute monocytic leukemia cell line treated with 5 ng/mL PMA for 48 hours and 100 ng/mL LPS and 3 µM monensin for 24 hours (positive staining) and THP‑1 human acute monocytic leukemia cell line (untreated, negative staining) using Mouse Anti-Human IL‑12 p70 Monoclonal Antibody (Catalog # MAB611R) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.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 p70 Antibody (24945R)
Application
Recommended Usage
Immunocytochemistry
8-25 µg/mL
Sample: IL‑12 p70 was detected in immersion fixed THP‑1 human acute monocytic leukemia cell line treated with 5 ng/mL PMA for 48 hours and 100 ng/mL LPS and 3 µM monensin for 24 hours
Sample: IL‑12 p70 was detected in immersion fixed THP‑1 human acute monocytic leukemia cell line treated with 5 ng/mL PMA for 48 hours and 100 ng/mL LPS and 3 µM monensin for 24 hours
Human IL-12 p70 Sandwich Immunoassay
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Formulation, Preparation, and Storage
Purification
Protein A or G purified from cell culture supernatant
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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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
Background: IL-12 p70
Long Name
Interleukin 12 p70
Alternate Names
IL12 p70
Gene Symbol
IL12A
UniProt
Additional IL-12 p70 Products
Product Documents for Human IL-12 p70 Antibody (24945R)
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human IL-12 p70 Antibody (24945R)
For research use only
Citations for Human IL-12 p70 Antibody (24945R)
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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