Product Information

Contents: Functional Grade Purified anti-mouse IL-23 (Interleukin-23) p19
Catalog Number: 16-7232
Sizes: 50 ug, 500 ug
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, No NaN₃
Storage Conditions: Store at 4°C.
Avoid repeated freeze/thaw cycles.
KEEP CONTENTS STERILE.
Clone: G23-8
Isotype: Rat IgG1, κ

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-7232	Affinity Purified anti-mouse IL-23 (Interleukin-23, IL23) p19	N/A	N/A	ELISA cap WB
16-7232	Functional Grade* Purified anti-mouse IL-23 (Interleukin-23, IL23) p19	N/A	N/A	ELISA cap NU WB
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.

Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq.

Description

The G23-8 antibody reacts with the p19 subunit of mouse IL-23. The G23-8 antibody was generated from immunization with authentic, insect cell-expressed, recombinant mouse IL-23 heterodimer. The G23-8 antibody can specifically neutralize IL-23 bioactivity with no effect on IL-12 p70 bioactivity.

The use of a p19-specific capture antibody and a p40-specific detection antibody yields an IL-23 ELISA which is exquisitely specific for mouse IL-23. IL-12 p40 homodimer and IL-12 p70 were each run in the assay at 500 ng/ml with no interference or cross-reactivity observed. A panel of 20 unrelated cytokines was run in the IL-23 ELISA at 100 ng/ml with no cross reactivity observed; all values were at the limit of detection of the assay. For measurement of total p40 protein levels, the Mouse IL-12/23 Total p40 ELISA Ready-SET-Go! is available (88-7120).

IL-23 is a heterodimeric cytokine composed of the p40 subunit of IL-12 disulfide-linked with a protein p19. p19, like p35 of IL-12, is biologically inactive by itself. IL-23 interacts with IL-12Rbeta1 and an additional, novel beta2-like receptor subunit with STAT4 binding domain, termed IL-23R. IL-23 is secreted by activated mouse and human dendritic cells. Biological activities of mouse IL-23 are distinct from those of mouse IL-12. Mouse IL-23 was found not to induce significant amounts of IFN-g. Mouse IL-23 does induce strong proliferation of memory T cells (but not naïve T cells), whereas IL-12 has no effect on memory cells. Additionally, mouse IL-23 (but not IL-12) can activate mouse memory T cells to produce the proinflammatory cytokine IL-17. Human IL-23 has biological properties which are less distinct from human IL-12; human IL-23 induces proliferation of memory T cells and induces moderate levels of IFN-g production by naïve and memory T cells, as compared to IL-12.

Applications Reported

For research use only, not for diagnostic or therapeutic use. The G23-8 antibody has been reported for use as the capture antibody in mouse IL-23 ELISA, for Western blotting, and for neutralization of mouse IL-23 bioactivity.

Applications Tested

The Functional Grade purified format of the G23-8 antibody has been tested as the capture antibody in an IL-23 ELISA assay (in conjunction with the biotinylated C17.8 antibody) and for neutralization of IL-23 biological activity.

For in vitro neutralization, the Functional Grade purified G23-8 antibody at 1.25 ug/ml has been found to neutralize by 50% the biological effect of 0.1 ng/ml mouse IL-23 in an assay of IL-17 secretion by mouse splenocytes. The G23-8 antibody has no effect on the bioactivity of mouse IL-12 p70. For neutralization of both IL-12 p70 and IL-23 p19p40, the anti-IL-12/IL-23 p40 antibody, clone C17.8, is recommended. Detailed information and protocols about cytokine bioassays and in vitro cytokine neutralization using antibodies can be found in the BestProtocols section.

A suitable range of concentrations of this antibody for ELISA capture is 1-4 µg/ml. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 4000 pg/ml - 30 pg/ml should be included in each ELISA plate.
Note: TMB, rather than ABTS, should be used as a substrate to achieve this sensitivity level.

References

Happel, K.I., et al. 2005. Divergent roles of IL-23 and IL-12 in host defense against Klebsiella pneumoniae. J. Exp. Med. 202: 761 - 769. [Mouse IL-23 ELISA]

Kidoya, H., et al. 2005. Fas ligand induces cell-autonomous IL-23 production in dendritic cells, a mechanism for fas ligand-induced IL-17 production. J. Immunol. 275: 8024-8031. [Mouse IL-23 ELISA]

Zakharova, M. et al. 2005. Paradoxical Anti-Inflammatory Actions of TNF- : Inhibition of IL-12 and IL-23 via TNF Receptor 1 in Macrophages and Dendritic Cells J. Immunol. 175: 5024 - 5033. [Mouse IL-23 ELISA]

Fichtner-Feigl, S., et al. 2005. Treatment of murine Th1- and Th2-mediated inflammatory bowel disease with NF-B decoy oligonucleotides. J. Clin. Invest. 115: 3057 - 3071. [Mouse IL-23 ELISA]

Kuwashima, N., et al. 2005. Delivery of Dendritic Cells Engineered to Secrete IFN- into Central Nervous System Tumors Enhances the Efficacy of Peripheral Tumor Cell Vaccines: Dependence on Apoptotic Pathways. J. Immunol. 175: 2730 - 2740. [Mouse IL-23 ELISA]

Hegazi, R.A.F., et al. 2005. Carbon monoxide ameliorates chronic murine colitis through a heme oxygenase 1–dependent pathway. J. Exp. Med. 202: 1703 - 1713. [Mouse IL-23 ELISA]

Ivanov, S., et al. 2007. Functional relevance of the IL-23-IL-17 axis in lungs in vivo. Am. J. Respir. Cell Mol. Biol. 36: 442-451. [Western blotting]

Leibundut-Landmann S, Grob O, et al. 2007. Syk- and CARD9-dependent coupling of innate immunity to the induction of T helper cells that produce interleukin 17. Nat Immunol. 2007 Jun;8(6):630-8. (FA, PubMed)

Sarkar S, Tesmer LA, et al. 2007. Interleukin-17 as a molecular target in immune-mediated arthritis: immunoregulatory properties of genetically modified murine dendritic cells that secrete interleukin-4. Arthritis Rheum. 56(1):89-100. (FA, PubMed)

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