Canine IL-12/IL-23 p40 Antibody

Catalog # Availability Size / Price Qty
AF1969
AF1969-SP
Cell Proliferation Induced by IL‑12 and Neutralization by Canine IL‑12/IL‑23 p40 Antibody.
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Canine IL-12/IL-23 p40 Antibody Summary

Species Reactivity
Canine
Specificity
Detects canine IL‑12/IL‑23 p40 in direct ELISAs and Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant canine IL‑12/IL‑23 p40
Ile23-Ser329
Accession # Q28268
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied 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 Canine IL‑12/IL‑23 p40 (Catalog # 1969-CL)
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 0.1-0.4 µg/mL in the presence of 25 ng/mL Recombinant Canine 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.

Data Example

Neutralization Cell Proliferation Induced by IL‑12 and Neutralization by Canine IL‑12/IL‑23 p40 Antibody. View Larger

Cell Proliferation Induced by IL‑12 and Neutralization by Canine IL‑12/IL‑23 p40 Antibody. Recombinant Canine IL‑12 (Catalog # 2118-CL) stimulates proliferation in PHA-activated human peripheral blood mononuclear cells (PBMC) in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Canine IL‑12 (25 ng/mL) is neutralized (green line) by increasing concen­trations of Goat Anti-Canine IL‑12/IL‑23 p40 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1969). The ND50 is typically 0.1-0.4 µg/mL.

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.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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/IL-23 p40

Interleukin 12 (IL-12) and IL-23 are secreted heterodimeric glycoproteins belonging to the IL-12 cytokine family. The two cytokines share a common p40 (40 kDa) subunit, which is disulfide-linked with the p35 (35 kDa) subunit in IL-12, and with the p19 (19 kDa) subunit in IL-23. Canine p40 is synthesized as a 329 amino acid (aa) precursor with a 22 aa signal sequence and a 307 aa mature region. It contains a 90 aa fibronectin type III domain and a 75 aa Ig C2-like region. The expression of p40 is induced by substances such as LPS and CpG that activate antigen-presenting cells. Besides being found as a component of IL-12 or IL-23, free p40 monomers and homodimers are also secreted by cells expressing p40. Canine p40 shares 94%, 85%, 84%, 65%, and 65% aa sequence identity with feline, human, porcine, rat and mouse p40, respectively. Cells known to express p40 include macrophages, dendritic cells, monocytes, Langerhans cells, neutrophils, keratinocytes, plasmacytoid dendritic cells, and microglia. From cells that express both the p35 and p40 subunits (dendritic cells, monocytes, and CHO cells), the amount of free p40 secreted is 10‑1000 fold more than the heterdimeric IL-12. The high-affinity IL-12 receptor complex that transduces IL-12 signals is composed of a 100 kDa ligand-binding subunit (IL-12 R beta 1) and a 130 kDa signal transducing subunit (IL-12 R beta 2). Similarly, the high-affinity IL-23 signaling receptor complex is composed of the shared IL-12 R beta 1 and the unique IL-23 R, a novel gp130-like protein. Both the monomeric and the dimeric free p40 can bind to the IL-12 R beta 1 and function as antagonists of IL-12 or IL-23. However, the monomeric p40 binds IL-12 R beta 1 with lower affinity and is less potent as an IL-12 antagonist. Homodimeric mouse p40 has also been shown to have agonistic functions similar to IL-12, inducing nitric oxide expression and NF kappa B activation in mouse primary microglia and peritoneal macrophages. The molecular mechanism for the agonistic effects of homodimeric p40 has not been determined (1‑6).

References
  1. Buttner, M. et al. (1997) Cytokine 10:241. 
  2. Park, A.Y. and P. Scott (2001) Scand. J. Immunol. 53:529.  
  3. Trinchieri, G. et al. (2003) Immunity 19:641.
  4. Brombacher, F. et al. (2003) Trends Immunol. 24:207.
  5. Lankford, C.S. and D.M. Frucht (2003) J. Leukoc. Biol. 73:49.
  6. Abdi, K. (2002) Scand. J. Immunol. 5:1.
Long Name
Interleukin 12/Interleukin 23 p40
Entrez Gene IDs
3593 (Human); 16160 (Mouse); 64546 (Rat); 403976 (Canine); 102124858 (Cynomolgus Monkey); 493741 (Feline); 694747 (Rhesus Macaque)
Alternate Names
CLMF p40; CLMF; CLMF2; Cytotoxic lymphocyte maturation factor 40 kDa subunit; IL12 p40; IL-12 p40; IL-12 subunit p40; IL12B; IL-12B; IL-12BNK cell stimulatory factor chain 2; interleukin 12, p40; interleukin 12B (natural killer cell stimulatory factor 2, cytotoxic lymphocytematuration factor 2, p40); interleukin-12 beta chain; interleukin-12 subunit beta; natural killer cell stimulatory factor, 40 kD subunit; NKSF; NKSF2; NKSF2IL12, subunit p40

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