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BD Pharmingen™ Biotin Mouse Anti-Human IL-12 (p40/p70)
Clone C8.6 (RUO)
Human IL-12 p70 Protein Quantitation by ELISA. Rat anti-human IL-12 p70 specific antibody 20C2 (Cat. No. 555065) was paired with a biotinylated anti-IL-12 p40/p70 detection (Cat. No. 554660) to measure specifically human IL-12 p70 protein, using recombinant human IL-12 p70 (Cat. No. 554613), as the ELISA standard.
Human IL-12 p70 Protein Quantitation by ELISA. Rat anti-human IL-12 p70 specific antibody 20C2 (Cat. No. 555065) was paired with a biotinylated anti-IL-12 p40/p70 detection (Cat. No. 554660) to measure specifically human IL-12 p70 protein, using recombinant human IL-12 p70 (Cat. No. 554613), as the ELISA standard.
Product Details
BD Pharmingen™
Human (QC Testing)
Mouse IgG1, κ
Purified p40 Subunit of Human IL-12
ELISA Detection (Routinely Tested)
0.5 mg/ml
AB_395495
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures
ELISA Detection (IL-12 p70 protein ELISA): The biotinylated C8.6 antibody (Cat. No. 554660) is useful as a detection antibody for a sandwich ELISA for specifically measuring human IL-12 p70 protein levels. Biotinylated C8.6 antibody can be paired with the purified rat anti-human IL-12 p70 specific 20C2 (Cat. No. 555065) as the capture antibody and recombinant human IL-12 p70 protein (Cat. No. 554613) as the standard. Biotinylated C8.6 antibody should be titrated (0.5 - 2.0 µg/ml) to determine optimal concentration for ELISA detection. To obtain linear standard curves, doubling dilutions of human IL-12 p70 ranging from ~2000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For testing IL-12 p70 in serum or plasma BD OptEIA™ Set (Cat. No. 555183) or BD OptEIA™ Kit (Cat. No. 559258) is recommended.
Note 1: This ELISA for measuring human IL-12 p70 protein levels is not interfered with by a 50-fold excess (by mass) of the human p40 monomer. This assay is specific for measuring human IL-12 p70 protein. It has been reported that activated PBMC produce an excess of free p40 protein over the p70 heterodimer.
ELISA Detection (IL-12 p40 protein ELISA): The biotinylated C8.6 antibody (Cat. No. 554660) is useful as a detection antibody for specifically measuring human IL-12 p40 protein levels, with no reactivity to human IL-12 p70 protein. Biotinylated C8.6 antibody can be paired with the purified (exclusively p40 specific) C8.3 antibody (Cat. No. 551227) as the capture antibody with recombinant human IL-12 p40 (Cat. No. 554633) as the standard. For testing IL-12 (p40) in complex biological fluids like serum or plasma BD OptEIA™ Set (Cat. No. 555171) or BD OptEIA™ Kit (Cat. No. 551116) is recommended.
Note 2: These ELISA pairs show no cross-reactivity with any of the cytokines or chemokines tested (e.g., human IL-1α, IL-1β, IL-2, IL-3, IL-4. IL-5, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-13, IL-15, G-CSF, GM-CSF, IFN-γ, lymphotactin, MCP-1, MCP-2, MIP-1α, MIP-1β, NT-3, PDGF-AA, sCD23, SCF, TNF, Lt-α, VEGF; mouse IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 p70, IL-15, GM-CSF, IFN-γ, MCP-1, TCA-3, TNF; rat IL-2, IL-4, IL-6, IL-10, GM-CSF, IFN-γ, TNF).
Neutralization: The C8.6 antibody has been reported to be useful for neutralization of human IL-12 p70 bioactivity. A formulation that does not contain sodium azide formulation (NA/LE™) is available (Cat. No. 554659) and is recommended for functional assay. This application is not routinely tested at BD Biosciences Pharmingen.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
Companion Products
Purified Rat Anti-Human IL-12 p70 RUO
Size
0.5 mg
Cat No.
555065
Recombinant Human IL-12 (p70) RUO
Size
5 µg
Cat No.
554613
Human IL-12 (p70) ELISA Set RUO
Size
20 Plate(s)
Cat No.
555183
Human IL-12 (p70) ELISA Kit RUO
Size
2 Plate(s)
Cat No.
559258
Purified Mouse Anti-Human IL-12 (p40) RUO
Size
1 mg
Cat No.
551227
Recombinant Human IL-12 p40 RUO
Size
5 µg
Cat No.
554633
554660 Rev. 1
Antibody Details
C8.6
The C8.6 monoclonal antibody specifically recognizes human IL-12 p40 that is encoded by IL12B (interleukin 12B). The antibody binds to IL-12 p40 monomers and homodimers as well as to the p40 subunit of the heterodimeric cytokines, IL-12 p70 (p35/p40) and IL-23 (p19/p40). These cytokines are produced by a variety of cells, including activated monocytes, macrophages, neutrophils, and dendritic cells. IL-12 p40 monomers and homodimers can block the binding of heterodimeric IL-12 to its receptor. IL-12 promotes the generation of Th1 cells and activates NK cells, Group 1 innate lymphoid cells (ILC1), and cytotoxic T cells leading to production of IFN-γ, TNF, granzymes, and perforin. IL-23 promotes the generation and activation of Th17 cells and other effector cells, including γδ T cells, NKT cells, MAIT cells, and ILC3 that can produce cytokines including IL-17A, IL-17F, IL-22, GM-CSF, TNF, and IL-6. The immunogen used to generate the C8.6 hybridoma was the purified p40 subunit of human IL-12. The C8.6 antibody is a neutralizing antibody and can reportedly inhibit the biological activities of IL-12 and IL-23.
This antibody is routinely tested by ELISA as a detection antibody. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.
554660 Rev. 1
Format Details
Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
554660 Rev.1
Citations & References
View product citations for antibody "554660" on CiteAb
Development References (3)
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D'Andrea A, Aste-Amezaga M, Valiante NM, Ma X, Kubin M, Trinchieri G. Interleukin 10 (IL-10) inhibits human lymphocyte interferon gamma-production by suppressing natural killer cell stimulatory factor/IL-12 synthesis in accessory cells. J Exp Med. 1993; 178(3):1041-1048. (Clone-specific: Neutralization). View Reference
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D'Andrea A, Rengaraju M, Valiante NM, et al. Production of natural killer cell stimulatory factor (interleukin 12) by peripheral blood mononuclear cells. J Exp Med. 1992; 176(5):1387-1398. (Clone-specific). View Reference
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Oppmann B, Lesley R, Blom B, et al. Novel p19 protein engages IL-12p40 to form a cytokine, IL-23, with biological activities similar as well as distinct from IL-12.. Immunity. 2000; 13(5):715-25. (Clone-specific: Immunoprecipitation). View Reference
554660 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.