Human IFN-gamma Antibody

Catalog #: MAB285R Datasheet / COA / SDS

Recombinant Monoclonal Antibody

Catalog #	Availability	Size / Price	Qty
MAB285R-100
MAB285R-SP

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Human IFN-gamma Antibody Summary

Species Reactivity

Human

Specificity

Detects human IFN-gamma in direct ELISA.

Source

Recombinant Monoclonal Mouse IgG_2A Clone # 25718R

Purification

Protein A or G purified from cell culture supernatant

Immunogen

E. coli-derived recombinant human IFN-gamma
Gln24-Gln166
Accession # AAP20098.1

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or 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

Immunocytochemistry

8-25 µg/mL

See below

Neutralization

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 0.02-0.06 µg/mL in the presence of 1 ng/mL Recombinant Human 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.

Scientific Data

Immunocytochemistry

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IFN‑ gamma in Human PBMCs. IFN-gamma was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with PHA using Mouse Anti-Human IFN-gamma Monoclonal Antibody (Catalog # MAB285R) at 8 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.

Neutralization

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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 (1 ng/mL) is neutralized (green line) by increasing concentrations of Human IFN-gamma Monoclonal Antibody (Catalog # MAB285R). The ND₅₀ is typically 0.02-0.06 µg/mL.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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, IFNG), 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 molecular weight variably glycosylated subunits. Glycosylation of IFN-gamma at sites Asn25 and Asn97 is critical for protease resistance. 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 function 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. 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.

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