IFN-gamma Antibody

R&D Systems | Catalog # AF-585-NA

R&D Systems
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Mouse, Rat

Cited:

Mouse, Rat

Applications

Validated:

Western Blot, Neutralization, Immunocytochemistry

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Neutralization

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all IFN-gamma Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant rat IFN-gamma
Gln23-Cys156
Accession # P01581

Specificity

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

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

Scientific Data Images for IFN-gamma Antibody

IFN‑ gamma Inhibition of EMCV-induced Cytopathy and Neutralization by Mouse/Rat IFN‑ gamma Antibody.

IFN‑ gamma Inhibition of EMCV-induced Cytopathy and Neutralization by Mouse/Rat IFN‑ gamma Antibody.

Recombinant Rat IFN-gamma (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-? antibody in Rat Splenocytes by Immunocytochemistry (ICC).

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; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Rat IFN-gamma ELISA Standard Curve

Rat IFN-gamma ELISA Standard Curve

Recombinant Rat IFN‑ gamma (585-IF) was serially diluted and captured by Mouse Anti-Rat IFN‑ gamma Monoclonal Antibody (MAB5851) coated on a Clear Polystyrene Microplate (DY990). Goat Anti-Mouse/Rat IFN‑ gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-585-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (DY998).
Detection of IFN-gamma by Immunocytochemistry/ Immunofluorescence

Detection of IFN-gamma by Immunocytochemistry/ Immunofluorescence

The effects of Hyperoside on the imbalance of T cell differentiation in the kidney. Immunofluorescence staining shows the expressions of CD4+ T-cell associated cytokines, IFN-gamma [red, (A)], IL-17 [red, (B)], and Foxp3 [red, (C)] in kidney tissue of db/db mice treated with HPS. Quantitative analyses of IFN-gamma (D), IL-17 (E), and Foxp3 (F) expressions in kidney tissue were also performed (Original magnification: ×400, scale bar = 50 μm). Values are presented as the means ± SD. Differences between experimental groups were evaluated using ANOVA. (n = 5 per group, #P < 0.05 and ##P <0.01 vs db/m; **P < 0.01 vs db/db; ns, not significant). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34925317), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for IFN-gamma Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed rat splenocytes either untreated or treated with PMA and calcium ionomycin

Western Blot

0.1 µg/mL
Sample: Recombinant Rat IFN-gamma (Catalog # 585-IF)

Neutralization

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.

Reviewed Applications

Read 1 review rated 5 using AF-585-NA in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.<br/>*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 ), 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).

References

  1. Billiau, A. and P. Matthys (2009) Cytokine Growth Factor Rev. 20:97.
  2. Pestka, S. et al. (2004) Immunol. Rev. 202:8.
  3. Dijkema, R. et al. (1985) EMBO J. 4:761.
  4. Marsters, S.A. et al. (1995) Proc. Natl. Acad. Sci. USA 92:5401.
  5. Krause, C.D. et al. (2000) J. Biol. Chem. 275:22995.
  6. Schroder, K. et al. (2004) J. Leukoc. Biol. 75:163.
  7. McLaren, J.E. and D.P. Ramji (2009) Cytokine Growth Factor Rev. 20:125.
  8. Muhl, H. and J. Pfeilschifter (2003) Int. Immunopharmacol. 3:1247.
  9. Kelchtermans, H. et al. (2008) Trends Immunol. 29:479.

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

UniProt

P01581

Additional IFN-gamma Products

Product Documents for IFN-gamma Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for IFN-gamma Antibody

For research use only

Citations for IFN-gamma Antibody

Customer Reviews for IFN-gamma Antibody (1)

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  • Mouse/Rat IFN-gamma Antibody
    Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: Mouse oral tumor cells
    Species: Mouse
    Verified Customer | Posted 08/21/2017
    Formalin fixed paraffin embedded mouse oral tumor specimens stained with IFN-gamma antibody at 1:3000 dilution.
    IFN-gamma Antibody AF-585-NA

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