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Anti mouse ifn γ mab

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The Anti-mouse-IFN-γ mAb is a monoclonal antibody that specifically binds to interferon-gamma (IFN-γ) of mouse origin. It is a laboratory research tool used to detect and quantify mouse IFN-γ in various biological samples.

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Lab products found in correlation

Elispot plate, by Merck Group (3 mentions) Streptavidin alkaline phosphatase, by Thermo Fisher Scientific (3 mentions) Series 2 immunospot analyzer, by Cellular Technology (3 mentions) Hl 1 medium, by Lonza (3 mentions) Anti mouse il 17 mab, by BioLegend (2 mentions) Anti mouse il17mab, by BioXCell (2 mentions) Siinfekl, by ProImmune (1 mentions)

Spelling variants of this product

IFN-γ (1815 citations) Anti-IFN-γ (147 citations) Mouse IFN-γ (48 citations) Anti-mouse IFN-γ (17 citations)

4 protocols using anti mouse ifn γ mab

1

Cytokine ELISPOT Assay for Antigen-Specific Responses

Cited 1 time
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Cytokine ELISPOT assay was performed and spots analyzed as described previously (18 (link), 19 (link)). In brief, ELISPOT plates (Millipore, Billerica, MA, USA) were precoated with anti-mouse-IFN-γ mAb (eBioscience, AN-18) and anti-mouse-IL-17 mAb (Bio X Cell; 17F3). Splenocytes (5 × 105 cells/well) and brain-infiltrating mononuclear cells (2–5 × 104 cells/well) were restimulated with MOG35–55 peptide in HL-1 medium (Lonza) at 37°C for 24 h. Biotinylated anti-mouse-IFN-γ mAb (eBioscience; R4-6A2) and anti-mouse-IL-17 mAb (BioLegend, TC11-8H4) were then added overnight at 4°C, followed by incubation with streptavidin alkaline phosphatase (Invitrogen, Waltham, MA, USA) for 2 h at room temperature and developing with BCIP/NBT Phosphatase Substrate (KPL, Gaithersburg, MD, USA). After plate developing, image analysis of spots was performed on a Series 2 Immunospot analyzer (Cellular Technology Limited, Cleveland, OH, USA). Results for antigen-specific spot-forming cells were normalized with a negative control containing peptide-free media. All measurements were performed in duplicate or triplicate.
Raphael I., Webb J., Gomez-Rivera F., Chase Huizar C.A., Gupta R., Arulanandam B.P., Wang Y., Haskins W.E, & Forsthuber T.G. (2017). Serum Neuroinflammatory Disease-Induced Central Nervous System Proteins Predict Clinical Onset of Experimental Autoimmune Encephalomyelitis. Frontiers in Immunology, 8, 812.
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2

Cytokine ELISPOT Assay for T-cell Analysis

Cited 1 time
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Cytokine ELISPOT assay was performed and spots analyzed as described previously.20 (link) In brief, ELISPOT plates (Millipore, Billerica, MA) were precoated with anti-mouse-IFN-γ mAb (eBioscience, San Diego, CA; AN-18) and anti-mouse-IL-17 mAb (Bio X Cell; 17F3). Splenocytes (1 × 106 cells per well) and brain-infiltrating mononuclear cells (MNCs; 2–5 × 104 cells per well) were restimulated with MOG35-55 peptide in HL-1 medium (Lonza, Basel, Switzerland) at 37°C for 24 hours. Biotinylated anti-mouse-IFN-γ mAb (eBioscience; R4-6A2) and anti-mouse-IL-17 mAb (BioLegend, San Diego, CA; TC11-8H4) were then added overnight at 4°C, followed by incubation with streptavidin alkaline phosphatase (Invitrogen, Waltham, MA) for 2 hours at room temperature and developing with BCIP/NBT Phosphatase Substrate (KPL, Gaithersburg, MD). After plate developing, image analysis of spots was performed on a Series 2 Immunospot analyzer (Cellular Technology Limited, Cleveland, OH).
Ji N., Kovalovsky A., Fingerle-Rowson G., Guentzel M.N, & Forsthuber T.G. (2015). Macrophage migration inhibitory factor promotes resistance to glucocorticoid treatment in EAE. Neurology® Neuroimmunology & Neuroinflammation, 2(5), e139.
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3

In Vivo Cytotoxicity Assay for CD8+ T Cells

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OVA-specific CD8+ T cells from SDLN or spleen were analyzed by MHC-class I pentamer (257–264 SIINFEKL, Proimmune) staining. CTL activity was determined in vivo as described (36 (link)). Briefly, syngeneic spleen cells were stained with 5 or 0.3 μM CFSE, respectively. The latter fraction was pulsed with 10 μM CTL peptide (OVA 257–264 SIINFEKL) (37 (link)) or βGal 96–103, DAPIYTNV) (38 (link)) and combined with the first (non-pulsed) fraction. Of this mixture, 3 × 106 cells were injected i.v. into recipient mice. After 16 hrs, splenocytes and SDLN cells were analyzed by flow cytometry, and % specific lysis was calculated from the reduction of peptide-pulsed CFSEdim target cells relative to non-pulsed CFSEhi reference cells. Antigen-specific IFNγ-secreting CD8+ T cells were detected by ELISPOT technique, as described elsewhere (33 (link)). In brief, whole splenocytes or LN cells were cultured on ELISPOT plates coated with anti-mouse IFNγ mAb (clone: AN-18, eBioscience) in the presence or absence of CTL peptide (SIINFEKL or DAPIYTNV). After 24 hrs, cytokine spots were detected by use of biotinylated anti-mouse IFNγ mAb (clone: R46A2; Biolegend), followed by streptavidin-HRP and 3-amino-9-ethylcarbazol as a chromogenic substrate. Spots were counted manually from flatbed scans of duplicate wells.
Strandt H., Pinheiro D.F., Kaplan D.H., Wirth D., Gratz I.K., Hammerl P., Thalhamer J, & Stoecklinger A. (2017). Neoantigen Expression in Steady-State Langerhans Cells Induces CTL Tolerance. The Journal of Immunology Author Choice, 199(5), 1626-1634.
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4

Cytokine ELISPOT for Antigen-Specific Responses

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Cytokine ELISPOT assay was performed and spots were analyzed as described previously [18 (link),19 (link)]. In brief, ELISPOT plates (Millipore, Billerica, MA, USA) were precoated with anti-mouse-IFN-γ mAb (eBioscience, San Diego, CA, USA, AN-18) and anti-mouse-IL-17 mAb (17F3, Bio X Cell, Lebanon, NH, USA). Splenocytes (5 × 105 cells/well) were restimulated with MOG35–55 peptide in HL-1 medium (Lonza, Basel, Switzaerland) at 37 °C, 5% CO2 for 24 h. Biotinylated anti-mouse-IFN-γ mAb (R4-6A2, eBioscience/Thermo Fisher, Waltham, MA, USA) and anti-mouse-IL-17 mAb (TC11-8H4, BioLegend, San Diego, CA, USA) were then added overnight at 4 °C, followed by incubation with streptavidin alkaline phosphatase (Invitrogen, Waltham, MA, USA) for 2 h at room temperature and developing with BCIP/NBT Phosphatase Substrate (KPL, Gaithersburg, MD, USA). After plate development, image analysis of spots was performed on a Series 2 ImmunoSpot analyzer (Cellular Technology Limited, Cleveland, OH, USA). Results for antigen-specific spot-forming cells were normalized with a negative control containing peptide-free media. All measurements were performed in triplicate.
Asmis R., Medrano M.T., Chase Huizar C., Griffith W.P, & Forsthuber T.G. (2024). Dietary Supplementation with 23-Hydroxy Ursolic Acid Reduces the Severity and Incidence of Acute Experimental Autoimmune Encephalomyelitis (EAE) in a Murine Model of Multiple Sclerosis. Nutrients, 16(3), 348.
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