Human IFN-gamma Antibody

Catalog # MAB285 | R&D Systems, Inc. a Bio-Techne Brand
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Key Product Details

Species Reactivity

Human

Applications

Immunocytochemistry, Intracellular Staining by Flow Cytometry, Neutralization

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 25718

Product Summary for Human IFN-gamma Antibody

Immunogen

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

Specificity

Detects human IFN-gamma.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human IFN-gamma Antibody

IFN‑? antibody in Human PBMCs by Immunocytochemistry (ICC).

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 Human IFN-gamma Monoclonal Antibody (Catalog # MAB285) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of IFN- gamma antibody in Human PBMCs antibody by Flow Cytometry.

Detection of IFN‑ gamma in Human PBMCs by Flow Cytometry.

Human peripheral blood mononuclear cells (PBMCs) treated with 50 ng/mL PMA, 1 ug/mL Ionomycin, and 3 uM Monensin overnight were stained with either (A) Mouse Anti-Human IFN- gamma Monoclonal Antibody (Catalog # MAB285) or (B) Mouse IgG2A Isotype Control (Catalog # MAB003) followed by anti-Mouse IgG PE-conjugated secondary antibody (Catalog # F0102B) and Mouse Anti-Human CD3 epsilon APC-conjugated Monoclonal Antibody (Catalog # FAB100A). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
IFN-gamma Inhibition of EMCV-induced Cytopathy and Neutralization by Human IFN-gamma Antibody.

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 # MAB285). The ND50 is typically 0.02-0.06 µg/mL.

Applications for Human IFN-gamma Antibody

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with PMA and ionomycin

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: Human PBMCs treated with PMA, Ionomycin, and Monensin fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012)

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 of this antibody should be experimentally determined.

Formulation, Preparation and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Reconstitution Buffer Available:
Size / Price
Qty

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.

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, 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

Alternate Names

IFG, IFI, IFNG, IFNgamma

Entrez Gene IDs

281237 (Bovine); 403801 (Canine); 493965 (Feline); 3458 (Human); 15978 (Mouse); 396991 (Porcine); 25712 (Rat)

Gene Symbol

IFNG

UniProt

AAP20098.1

Product Documents for Human IFN-gamma Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human IFN-gamma Antibody

For research use only

Citations for Human IFN-gamma Antibody (25)

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Customer Reviews for Human IFN-gamma Antibody (4)

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  • Human IFN-gamma Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Peripheral blood mononuclear cells (PBMCs)
    Species: Human
    Verified Customer | Posted 06/17/2022
    Human IFN-gamma Antibody MAB285
  • Human IFN-gamma Antibody
    Name: Anonymous
    Application: Block/Neutralize
    Sample Tested: Cell Culture Media
    Species: Human
    Verified Customer | Posted 04/28/2021
    Human IFN-gamma Antibody MAB285
  • Human IFN-gamma Antibody
    Name: Anonymous
    Application: Microarrays
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 03/11/2019
    Human IFN-gamma Antibody MAB285
  • Human IFN-gamma Antibody
    Name: Carly Garrison
    Application: Microarray
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 01/09/2019

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