Bovine IFN-gamma Antibody

  
  • Species Reactivity
    Bovine
  • Specificity
    Detects bovine IFN-gamma in direct ELISAs and Western blots. In Western blots, 100% cross-reactivity with IFN-gamma from equine, canine, or feline systems is observed and no cross-reactivity with human, cotton rat, mouse, porcine, or rat IFN-gamma is observed.
  • Source
    Monoclonal Rat IgG2A Clone # 345025
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant bovine IFN-gamma
    Gln24-Thr166
    Accession # NP_776511
  • 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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    Recombinant Bovine IFN-gamma (Catalog # 2300-BG)
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    2-10 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    Bovine peripheral blood mononuclear cells treated with PMA and Calcium Ionomycin, fixed with paraformaldehyde, and permeabilized with saponin
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
Immunocytochemistry
IFN‑ gamma in Bovine PBMCs. IFN‑ gamma was detected in immersion fixed bovine peripheral blood mononuclear cells (PBMCs) treated with Calcium Ionomycin and PMA using Rat Anti-Bovine IFN‑ gamma Monoclonal Antibody (Catalog # MAB2300) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 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 bovine IFN-gamma exists as a noncovalently linked homodimer of 20‑25 kDa variably glycosylated subunits (3). It shares 78%‑80% amino acid (aa) sequence identity with canine, feline, equine, and porcine IFN-gamma and 42%‑59% with cotton rat, human, mouse, rat, and rhesus 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. Cerretti, D.P. et al. (1986) J. Immunol. 136:4561.
    4. Marsters, S.A. et al. (1995) Proc. Natl. Acad. Sci. 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
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