Detects rat and mouse IFN-gamma in direct ELISAs and Western blots. In direct ELISAs, approximately 20% cross-reactivity with recombinant human (rh) IFN-gamma, recombinant canine IFN-gamma, recombinant feline IFN‑ gamma, recombinant equine IFN-gamma, recombinant porcine IFN-gamma, recombinant rhesus monkey IFN-gamma, recombinant cotton rat IFN-gamma, and recombinant bovine IFN‑ gamma is observed. In Western blots, less than 10% cross‑reactivity with rhIFN‑ gamma is observed.
Polyclonal Goat IgG
E. coli-derived recombinant rat IFN-gamma Gln23-Cys156 Accession # P01581
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.
<0.10 EU per 1 μg of the antibody by the LAL method.
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 0.08-0.2 µg/mL in the presence of 2.5 ng/mL Recombinant Rat 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.
IFN‑ gamma Inhibition of EMCV-induced Cytopathy and Neutralization by Mouse/Rat IFN‑ gamma Antibody. Recombinant Rat IFN‑ gamma (Catalog # 585-IF) 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 crystal violet staining. Inhibition of EMCV activity elicited by Recombinant Rat IFN‑ gamma (2.5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Rat IFN‑ gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-585-NA). The ND50 is typically 0.08‑0.2 µg/mL.
IFN‑ gamma in Rat Splenocytes. IFN‑ gamma was detected in immersion fixed rat splenocytes treated with PMA and calcium ionomycin using Goat Anti-Mouse/Rat IFN‑ gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-585-NA) 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 cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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.
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 rat IFN-gamma exists as a noncovalently linked homodimer of 20-25 kDa variably glycosylated subunits (3). It shares 86% amino acid sequence identity with mouse IFN-gamma and 37-45% 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 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).
Billiau, A. and P. Matthys (2009) Cytokine Growth Factor Rev. 20:97.
Pestka, S. et al. (2004) Immunol. Rev. 202:8.
Dijkema, R. et al. (1985) EMBO J. 4:761.
Marsters, S.A. et al. (1995) Proc. Natl. Acad. Sci. USA 92:5401.
Krause, C.D. et al. (2000) J. Biol. Chem. 275:22995.
Schroder, K. et al. (2004) J. Leukoc. Biol. 75:163.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
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.