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Human IFN-gamma Antibody (25718)

R&D Systems | Catalog # MAB285

R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Neutralization, Intracellular Staining by Flow Cytometry, Immunocytochemistry

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Neutralization, Flow Cytometry, Bioassay, Differentiation, ELISA Detection, Functional Assay, Luminex (Detection), Stimulation

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 25718
View all IFN-gamma Primary Antibodies »

Product Specifications

Immunogen

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

Specificity

Detects human IFN-gamma.

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 (25718)

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.
Detection of IFN-gamma by Immunohistochemistry

Detection of IFN-gamma by Immunohistochemistry

IFN gamma and cytokine mix stimulation enhance epithelial barrier formation, brush border enzyme activity and disappearance of supranuclear vacuoles in the 2D intestinal monolayer(A) TEER values for organoids monolayers upon IFN gamma and cytokine mix stimulation compared to control (n = 12 independent donors).(B–D) (B) Alkaline phosphatase (μg pNPP/min, n = 8 independent donors), (C) LCT (μg glucose/min converted, n = 5 independent donors), and (D) SIS activity levels (μg glucose/min converted, n = 5 independent donors), measured on organoids 2D monolayers upon IFN gamma and cytokine mix stimulation compared to control.(E) Alkaline phosphatase staining showing 2D monolayer in embedded transwell upon IFN gamma and cytokine mix stimulation compared to control.(F) Transmission electron microscopy (TEM) shows loss of vacuoles (red arrows) upon IFN gamma, and cytokine mix stimulation compared to control. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, as determined by non-parametric one-way ANOVA. Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S2589004224011313), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of IFN-gamma by Immunohistochemistry

Detection of IFN-gamma by Immunohistochemistry

Development of alopecia areata (AA) in normal human scalp skin xenotransplants treated with (A) ILC1lc.Infiltrates of IFN-gamma +, CD8, and CD4 cells in group treated with (B) enriched CD8/NKG2D cells. Decreased number of IFN-gamma +, CD8, and CD4 cells in the (C) PBMCs/PHA treated one. (D) Quantitative data (E) presence of ILC1lc in transplants injected with ILC1lc compared to the virtual absence of these cells in enriched CD8/NKG2D and PBMCs/PHA treated mice.(F) Quantitative data N=5–9 xenotransplats/group from three donors. 4–5 areas were evaluated per section, and three sections per xenotransplant. Following Shapiro-Wilk test, Student’s t-test: *p<0.05, **p<0.01, ***p<0.001. Scale bar, 50 µm. DP - dermal papilla,HM - hair matrix.Figure 7—figure supplement 3—source data 1.Quantitative data for development of AA in normal human scalp skin xenotransplants treated with ILC1lc, enriched CD8/NKG2D and PBMCs/PHA.Quantitative data for development of AA in normal human scalp skin xenotransplants treated with ILC1lc, enriched CD8/NKG2D and PBMCs/PHA. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36930216), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of IFN-gamma by Immunohistochemistry

Detection of IFN-gamma by Immunohistochemistry

Impact of cytokines on metabolism and aggressiveness of SKOV3 cells. (A) TAG and TC NIR h2SRS imaging maps of SKOV3 cells with and without cytokines: IL‐2, TNF‐𝛼, IFN‐𝛾 supplemented media for SKOV3 cells, IL‐2 Concentration: 100 IU/mL, TNF‐𝛼 Concentration: 10 ng/mL, IFN‐𝛾 Concentration: 5ng/mL, scale bars: 10 µm. (B) Quantification of cellular TC and TAG in SKOV3 cells incubated with medium with and without cytokines supplementation, n>31 per group. (C) Confocal fluorescence imaging of migrated SKOV3 cells with and without cytokines, scale bars: 100 µm. (D) Quantification of the cell migration with and without Cytokines treatment, n = 3, ***p<0.001, **p<0.01, data presented as means +/± stdev, TC: total cholesterol. TAG: triglycerides/triacylglycerols. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/41691468), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of IFN-gamma by Flow Cytometry

Detection of IFN-gamma by Flow Cytometry

Transition of anagen to catagen hair follicles (HFs) following culture with ILC1lc or CD8+/NKG2D+ cells in human scalp HF ex vivo.(A) These immune cells significantly accelerated the transformation of anagen HFs into catagen HFs ex vivo compared to ILC2, ILC3, PBMCs/PHA, and neutralizing anti- IFN-gamma, anti-NKG2D antibodies. N=28–34 HFs/group taken from six independent donors, Student’s t-test: *p<0.05, **p<0.01, ***p<0.001. (B) ELISA analysis revealed increased IFN-gamma production by ILC1lc /HFs compared to production by CD8+/NKG2D+ cells, ILC2s, ILC3s, and PBMCs/PHA. N=6 healthy donors, 6 × 106 cells from each donor. Following Shapiro-Wilk test, Student’s t-test: *p<0.05, **p<0.01, ***p<0.001. (C) FACS analysis revealed a significant increased intracellular IFN-gamma expression in ILC1lc co-cultured with HFs compared to the effector CD8+/NKG2D+and to ILC2s and ILC3s, N=6 blood donors, 1.5 × 106 cells/blood donor. Student’s t-test, p<0.05.Figure 6—source data 1.Quantitative data for transition of anagen to catagen HFs following culture with ILC1lc or CD8+/NKG2D+ cells in human scalp HF ex vivo.Quantitative data for transition of anagen to catagen HFs following culture with ILC1lc or CD8+/NKG2D+ cells in human scalp HF ex vivo. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36930216), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of IFN-gamma by Immunohistochemistry

Detection of IFN-gamma by Immunohistochemistry

IFN gamma and cytokine mix stimulation increases IAP activity and FABP6 protein expression in HFOs(A and B) Alkaline phosphatase activity level (μg pNPP/min) in (A) proximal and (B) distal HFOs upon cytokines stimulation (n = 9 independent donors).(C) Alkaline phosphatase (IAP) staining of embedded HFOs upon IFN gamma, and cytokine mix stimulation compared to control.(D) FABP6 protein expression measured by western blot in distal HFOs.(E) FABP6 western blot quantification on n = 5 independent donors. ∗p < 0.05, ∗∗p < 0.01 as determined by non-parametric one-way ANOVA. Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S2589004224011313), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human IFN-gamma Antibody (25718)

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.

Reviewed Applications

Read 4 reviews rated 4.3 using MAB285 in the following applications:

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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 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

UniProt

AAP20098.1

Additional IFN-gamma Products

Product Documents for Human IFN-gamma Antibody (25718)

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Product Specific Notices for Human IFN-gamma Antibody (25718)

For research use only

Citations for Human IFN-gamma Antibody (25718)

Customer Reviews for Human IFN-gamma Antibody (25718) (4)

4.3 out of 5
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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 (25718) 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 (25718) 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 (25718) 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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