Measured by its ability to neutralize IFN‑ gamma inhibition of EMCV-induced cytopathy in the HeLa human cervical epithelial carcinoma cell line. Meager, A. (1987) in Lymphokines and Interferons, a Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 129. The Neutralization Dose (ND50) is typically 0.1-0.3 µg/mL in the presence of 5 ng/mL Recombinant Human IFN‑ gamma.
Please Note: Optimal dilutions should be determined by each laboratory for each application.
are available in the Technical Information section on our website.
IFN‑ gamma Inhibition of EMCV-induced Cytopathy and Neutralization by Human IFN‑ gamma Antibody.
Recombinant Human IFN‑ gamma (Catalog # 285-IF) reduces the Encephalomyocarditis Virus (EMCV)-induced cytopathy in the HeLa human cervical epithelial carcinoma cell line in a dose-dependent manner (orange line), as measured by crystal violet staining. Inhibition of EMCV activity elicited by Recombinant Human IFN‑ gamma (5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IFN‑ gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-285-NA). The ND50 is typically 0.1‑0.3 µg/mL.
IFN‑ gamma in Human PBMCs.
IFN‑ gamma was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with PMA and ionomycin using 10 µg/mL Goat Anti-Human IFN‑ gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # AF‑285‑NA) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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. Mature human IFN-gamma exists as a non-covalently linked homodimer of 20-25 kDa variably glycosylated subunits. It shares 90% amino acid (aa) sequence identity with rhesus IFN-gamma, 59-64% with bovine, canine, equine, feline, and porcine IFN‑ gamma, and 37-43% 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. IFN-gamma is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells. 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. In addition, IFN-gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation. The pleiotropic effects of IFN-gamma contribute to the development of multiple aspects of atherosclerosis.
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Immunohistochemistry: Human IFN-gamma Antibody [AF-285-NA].
Other Experimental Details
Other Experimental Details
Published in https://www.ncbi.nlm.nih.gov/pubmed/28169287Used at 10ug/ml.Briefly, frozen brain sections were fixed in 4% PFA (Fisher Scientific), followed by antigen retrieval using heating in acid citric buffer (Vector, Burlingame, CA, USA). Endogenous avidin-biotin was blocked for 15 min (Vector). Sections were incubated with 10% horse serum in PBS (Biosera, Boussens, France) and Fc Receptor Blocking Solution was added (Human TruStain FcX Biolegend,London, UK). Primary antibody was added overnight at 4 °C and detected with donkey anti-goat-biotin (ab6578, Abcam), followed by streptavidin-alkaline phosphatase (SA-5100, Vector) and visualised with the Vector Blue Alkaline Phosphatase Substrate Kit III (Vector).