Canine IFN-gamma Antibody

(2 citations)   
  • Species Reactivity
    Canine
  • Specificity
    Detects canine IFN-gamma in direct ELISAs and Western blots. In direct ELISAs, less than 40% cross-reactivity with recombinant porcine IFN‑ gamma is observed, approximately 20% cross-reactivity with recombinant feline IFN‑ gamma is observed, and less than 5% cross-reactivity with recombinant human IFN-gamma, recombinant rhesus macaque IFN-gamma, recombinant mouse IFN‑ gamma, and recombinant rat IFN‑ gamma is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant canine IFN-gamma
    Gln24-Lys166
    Accession # P42161
  • 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 IFN-gamma (Catalog # 781-CG)
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize IFN‑ gamma inhibition of VSV-induced cytopathy in the A‑72 canine fibroma cell line. The Neutralization Dose (ND50) is typically 0.2-1 µg/mL in the presence of 1.5 ng/mL Recombinant Canine IFN‑ gamma.
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 Examples

IFN‑ gamma Inhibition of VSV-induced Cytopathy and Neutralization by Canine IFN‑ gamma Antibody. Recombinant Canine IFN‑ gamma (Catalog # 781-CG) reduces the Vesticular Stomatitis Virus (VSV)-induced cytopathy in the A‑72 canine fibroma cell line in a dose-dependent manner (orange line), as measured by crystal violet staining. Inhibition of VSV activity elicited by Recombinant Canine IFN‑ gamma (1.5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Canine IFN‑ gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # AF781). The ND50 is typically 0.2-1 µg/mL.

Immunocytochemistry
IFN‑ gamma in Canine PBMCs. IFN‑ gamma was detected in immersion fixed canine peripheral blood mononuclear cells (PBMCs) untreated (lower panel) or treated with PMA and calcium ionomycin (upper panel) using Goat Anti-Canine IFN‑ gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # AF781) 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 cytoplasmic. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • 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: IFN-gamma
Interferon-gamma (IFN-gamma ), also known as type II or immune interferon, exerts a wide range of immunoregulatory activities and is considered to be the prototype proinflammatory cytokine (1, 2). Mature canine IFN-gamma exists as a noncovalently linked homodimer of 20‑25 kDa variably glycosylated subunits (3). It shares 79‑88% amino acid sequence identity with bovine, equine, and feline IFN-gamma, 62‑73% with human, porcine, and rhesus IFN-gamma, and 40‑47% with cotton rat, mouse, and rat IFN-gamma. IFN-gamma dimers bind to IFN-gamma RI (alpha subunits) which then interact with IFN-gamma RII (beta subunits) to form the functional receptor complex of two alpha and two beta subunits. Inclusion of IFN-gamma RII increases the binding affinity for ligand and the efficiency of signal transduction (4, 5). IFN-gamma is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells (6). It plays a key role in host defense by promoting the development and activation of Th1 cells, chemoattraction and activation of monocytes and macrophages, up regulation of antigen presentation molecules, and immunoglobulin class switching in B cells. It also exhibits antiviral, antiproliferative, and apoptotic effects (6, 7). In addition, IFN-gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation (8, 9). The pleiotropic effects of IFN-gamma contribute to the development of multiple aspects of atherosclerosis (7).
  • References:
    1. Billiau, A. and P. Matthys (2009) Cytokine Growth Factor Rev. 20:97.
    2. Pestka, S. et al. (2004) Immunol. Rev. 202:8.
    3. Zucker, K. et al. (1992) J. Interferon Res. 12:191.
    4. Marsters, S.A. et al. (1995) Proc. Natl. Acad. Sci. USA 92:5401.
    5. Krause, C.D. et al. (2000) J. Biol. Chem. 275:22995.
    6. Schroder, K. et al. (2004) J. Leukoc. Biol. 75:163.
    7. McLaren, J.E. and D.P. Ramji (2009) Cytokine Growth Factor Rev. 20:125.
    8. Muhl, H. and J. Pfeilschifter (2003) Int. Immunopharmacol. 3:1247.
    9. Kelchtermans, H. et al. (2008) Trends Immunol. 29:479.
  • Long Name:
    Interferon gamma
  • Entrez Gene IDs:
    3458 (Human); 15978 (Mouse); 25712 (Rat); 396991 (Porcine); 281237 (Bovine); 403801 (Canine); 493965 (Feline)
  • Alternate Names:
    IFG; IFI; IFNG; IFNgamma; IFN-gamma; Immune interferon; interferon gamma; interferon, gamma
Related Research Areas
Citations:

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2 Citations: Showing 1 - 2
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Species
Applications
Sample Type
  1. Mechanisms of Immune Suppression Utilized by Canine Adipose and Bone Marrow-Derived Mesenchymal Stem Cells
    Stem Cells Dev., 2016;0(0):.
    Species: Canine
    Sample Type: Whole Cells
    Application: Neut
  2. Impact of plasmid supercoiling on the efficacy of a rabies DNA vaccine to protect cats.
    Authors: Cupillard L, Juillard V, Latour S, Colombet G, Cachet N, Richard S, Blanchard S, Fischer L
    Vaccine, 2005;23(16):1910-6.
    Species: Feline
    Sample Type: Whole Cells
    Application: ELISpot Development
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