Human IFN-gamma Antibody (K3.53)

R&D Systems | Catalog # MAB2852

R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, ELISA Capture (Matched Antibody Pair), Neutralization, Intracellular Staining by Flow Cytometry

Cited:

Neutralization, Flow Cytometry, Array Development, ELISA Capture, ELISA Development, ELISA Development (Capture), Functional Assay, Luminex Development

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # K3.53
View all IFN-gamma Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human IFN-gamma

Specificity

Detects human IFN‑ gamma in direct ELISAs and Western blots. In Western blots, no cross-reactivity with recombinant mouse IFN‑ gamma, recombinant rat IFN‑ gamma, recombinant porcine IFN‑ gamma, recombinant feline IFN‑ gamma, or recombinant cotton rat IFN‑ gamma is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Endotoxin Level

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

Scientific Data Images for Human IFN-gamma Antibody (K3.53)

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 # MAB2852) 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-gammaamma (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-gammaamma (1 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IFN-gammaamma Monoclonal Antibody (Catalog # MAB2852). The ND50 is typically ≤ 2 µg/mL.

Applications for Human IFN-gamma Antibody (K3.53)

Application
Recommended Usage

Intracellular Staining by Flow Cytometry

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

Western Blot

1 µg/mL
Sample: Recombinant Human IFN-gamma (Catalog # 285-IF)

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  ≤2 µg/mL in the presence of 1 ng/mL Recombinant Human IFN‑ gamma.

Human IFN-gamma Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 2-8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human IFN‑ gamma Biotinylated Antibody (Catalog # BAF285)
  • Standard: Recombinant Human IFN-gamma Protein (Catalog # 285-IF)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 5 using MAB2852 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

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

Calculators

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.

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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

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

Gene Symbol

IFNG

Additional IFN-gamma Products

Product Documents for Human IFN-gamma Antibody (K3.53)

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human IFN-gamma Antibody (K3.53)

For research use only

Citations for Human IFN-gamma Antibody (K3.53)

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  • Human IFN-gamma Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HeLa human cervical epithelial carcinoma cell line
    Species: Human
    Verified Customer | Posted 08/24/2021
    Human IFN-gamma Antibody (K3.53) MAB2852

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