Mouse IFN-gamma Antibody

Recombinant Monoclonal Antibody.
  
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse IFN-gamma in Western blots. In Western blots, this antibody does not cross-react with recombinant human (rh) IFN‑ gamma, rrIFN‑ gamma, rpIFN‑ gamma, rrmIFN‑ gamma, rfeIFN‑ gamma, or rcaIFN‑ gamma.
  • Source
    Recombinant Monoclonal Rat IgG2A Clone # 37895R
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    E. coli-derived recombinant mouse IFN-gamma
  • Formulation
    Supplied as a solution in PBS. *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
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
  • Neutralization
    Measured by its ability to neutralize IFN‑ gamma inhibition of EMCV-induced cytopathy in the L‑929 mouse fibroblast cell line. Vogel, S. and M. Hogan (1995) in Current Protocols in Immunology. Ciocio, R. (ed); John Wiley & Sons, Inc. p. 6. 9. 1. The Neutralization Dose (ND50) is typically 0.075-0.3 µg/mL in the presence of 1.5 ng/mL Recombinant Mouse 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 EMCV-induced Cytopathy and Neutralization by Mouse IFN-gamma Antibody. Recombinant Mouse IFN-gamma (Catalog # 485-MI) reduces the Encephalomyocarditis Virus (EMCV)-induced cytopathy in the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by Resazurin (Catalog # AR002). Inhibition of EMCV activity elicited by Recombinant Mouse IFN-gamma (1.5 ng/mL) is neutralized (green line) by increasing concentrations of Mouse IFN-gamma Monoclonal Antibody (Catalog # MAB485R). The ND50 is typically 0.075-0.3 µg/mL.
Intracellular Staining by Flow Cytometry
Detection of IFN‑ gamma in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes were stained with Rat Anti-Mouse CD28 Alexa Fluor® 488‑conjugated Monoclonal Antibody (Catalog # FAB4832G) and either (A) Rat Anti-Mouse IFN‑ gamma Monoclonal Antibody (Catalog # MAB485R) or (B) Rat IgG2A Isotype Control (Catalog # MAB006) followed by Phycoerythrin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0105B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
  • Shipping
    The product is shipped with polar packs. 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 opening.
    • 6 months, -20 to -70 °C under sterile conditions after opening.
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 pro-inflammatory cytokine (1, 2). Mature mouse IFN-gamma exists as a noncovalently linked homodimer of 20-25 kDa variably glycosylated subunits (3). It shares 86% amino acid sequence identity with rat IFN-gamma and 38-44% with bovine, canine, cotton rat, equine, feline, human, porcine, and rhesus macaque 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, upregulation of antigen presentation molecules, and immunoglobulin class switching in B cells. It also exhibits anti-viral, anti-proliferative, 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. Gray, P.W. and D.V. Goeddel (1983) Proc. Natl. Acad. Sci. USA 80:5842.
    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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