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Mouse ifn γ duoset elisa kit

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The Mouse IFN-γ DuoSet ELISA kit is a laboratory reagent used to quantitatively measure mouse interferon-gamma (IFN-γ) levels in biological samples. It is a sandwich enzyme-linked immunosorbent assay (ELISA) that utilizes a capture and detection antibody pair specific for mouse IFN-γ.

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Lab products found in correlation

Mouse tnf α duoset elisa kit, by R&D Systems (3 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Red blood cell lysis buffer, by Merck Group (1 mentions) X rad 320 irradiator, by Precision X-Ray (1 mentions) Anti mouse cd28, by Thermo Fisher Scientific (1 mentions) Cd3 cd28 beads, by Thermo Fisher Scientific (1 mentions) Anti cd3 clone 145 2c11, by Thermo Fisher Scientific (1 mentions) Mouse cytokine array panel a, by R&D Systems (1 mentions) Ifn γ, by ProSpec (1 mentions) Ovalbumin (ova), by Thermo Fisher Scientific (1 mentions) Trp 2, by AnaSpec (1 mentions) Mouse ifnγ, by ProSpec (1 mentions) Radioimmunoprecipitation assay buffer, by Merck Group (1 mentions) Tissuelyser 2, by Qiagen (1 mentions) Cd3 clone 145 2c11, by BioLegend (1 mentions) Cd8a t cell isolation kit, by Miltenyi Biotec (1 mentions) 3 3 5 5 tetramethylbenzidine (tmb), by Tiangen Biotech (1 mentions) Enspire 2300 elisa reader, by PerkinElmer (1 mentions) Mouse il 1β, by Thermo Fisher Scientific (1 mentions) Mouse il 6 duoset elisa kit, by R&D Systems (1 mentions)

16 protocols using mouse ifn γ duoset elisa kit

1

T Cell Activation Assay for Immunotherapies

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Tumor cells were irradiated (5 Gy) with an X-RAD 320 irradiator (Precision X-Ray Inc.), and 1 × 104 tumor cells were seeded in a 96-well plate. Spleens from C57BL/6 mice were dissociated into a single-cell suspension using a 40-μm cell strainer, treated with red blood cell lysis buffer (Sigma-Aldrich, R7757), and further passed through a 100-μm cell strainer. A splenic single-cell suspension was then resuspended in complete IMDM (cIMDM) (Welgene, LM004-01) supplemented with 10% FBS, 1× anti-anti, and 55 μM 2-mercaptoethanol (Gibco, 21985). Splenocytes (5 × 105) were placed in a 96-well plate and cultured with In Vivo Ready anti-mouse CD3ε mAb (1 μg/ml) (clone: 145-2C11; Tonbo Biosciences, 40-0031) alone or with purified antibodies at the indicated dose. For cross-linking HER2×1D8, hIgG1 κ (1 μg/ml) (Southern Biotech, 0151K-01) was added. Seventy-two hours after culture, T cell activation was analyzed by flow cytometry, and supernatants were collected and assayed for IFN-γ secretion by ELISA using a mouse IFN-γ DuoSet ELISA kit (R&D Systems, DY485).
You G., Lee Y., Kang Y.W., Park H.W., Park K., Kim H., Kim Y.M., Kim S., Kim J.H., Moon D., Chung H., Son W., Jung U.J., Park E., Lee S., Son Y.G., Eom J., Won J., Park Y., Jung J, & Lee S.W. (2021). B7-H3×4-1BB bispecific antibody augments antitumor immunity by enhancing terminally differentiated CD8+ tumor-infiltrating lymphocytes. Science Advances, 7(3), eaax3160.
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2

Stimulation of CD8+ T Cells

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Purified CD8+ T cells were seeded into 96-well U-bottomed plates (105 cells/well) pre-coated with 5 μg/mL anti-CD3 (clone 145-2C11, eBioscience, San Diego, CA, USA). Anti-mouse CD28 (2 μg/mL) (clone 37.51, eBioscience), anti-mouse 4-1BB (5 μg/mL) (Leinco Technologies, St. Louis, MO, USA), and CD3/CD28 beads (Thermo Fisher Scientific, Waltham, MA, USA) were used as a positive control. 200 nM 3WJ only (control), 3WJ/4-1BB-M, 3WJ/4-1BB-D, 3WJ/4-1BB-T, and 3WJ/CD28-T were added to a final culture volume of 200 μL/well in complete RPMI 1640 medium and incubated at 37°C and 5% CO2 for 72 h. Afterward, 50 μL of cell culture supernatants was collected, and concentrations of IFN-γ were determined using a mouse IFN-γ DuoSet ELISA kit (R&D Systems, Minneapolis, MN, USA) following the protocol provided by the manufacturer. Data represent the mean ± standard deviation (SD) of three independent experiments.
Shu D., Zhang L., Bai X., Yu J, & Guo P. (2021). Stoichiometry of multi-specific immune checkpoint RNA Abs for T cell activation and tumor inhibition using ultra-stable RNA nanoparticles. Molecular Therapy. Nucleic Acids, 24, 426-435.
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3

Cytokine Profiling in Mice

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A mouse cytokine array panel A (ARY006, R&D Systems) was used for the mouse cytokine array according to the manufacturer’s instructions. The levels of CXCL1, CXCL2, TNF-α, and IFN-γ were measured using the mouse CXCL1 ELISA kit (LS-F268, LSBio, Seattle, Washington, USA), mouse CXCL2 ELISA kit (LS-F621, LSBio), mouse TNF-α DuoSet ELISA kit (DY-410, R&D Systems), and mouse IFN-γ DuoSet ELISA kit (DY-485, R&D Systems), respectively. All ELISAs were performed according to the manufacturers’ instructions.
Hwang Y.S., Cho H.J., Park E.S., Lim J., Yoon H.R., Kim J.T., Yoon S.R., Jung H., Choe Y.K., Kim Y.H., Lee C.H., Kwon Y.T., Kim B.Y, & Lee H.G. (2022). KLK6/PAR1 Axis Promotes Tumor Growth and Metastasis by Regulating Cross-Talk between Tumor Cells and Macrophages. Cells, 11(24), 4101.
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4

Evaluating Interferon-Gamma Response in B16F10 Cells

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For preparation of the target cells, B16F10 Red-FLuc cells were infected with the virus (100 PFU/cell). Four hours later, 50 units/ml mouse interferon gamma (IFNγ; Prospec Protein Specialists) was added to the culture. Forty-eight hours after viral infection, the cells were fixed with 1% paraformaldehyde and washed before added to the co-culture. For immune cell stimulation, pre-fixed target cells (2 × 104 cells/well) were incubated with splenocytes or lymph node cells (5 × 105 cells/well). Forty hours after the co-culture in a round-bottom 96-well plate, the concentration of IFNγ in the medium was assessed with a Mouse IFNγ DuoSet ELISA kit (R&D Systems).
Jiang H., Shin D.H., Nguyen T.T., Fueyo J., Fan X., Henry V., Carrillo C.C., Yi Y., Alonso M.M., Collier T.L., Yuan Y., Lang F.F, & Gomez-Manzano C. (2019). Localized Treatment with Oncolytic Adenovirus Delta-24-RGDOX Induces Systemic Immunity against Disseminated Subcutaneous and Intracranial Melanomas. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(22), 6801-6814.
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5

Evaluating IFNγ-Induced Antigen Presentation

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To prepare the target cells, 50 units/mL mouse IFNγ (Prospec Protein Specialists) was added to the B16-OVA cell culture. Forty-eight hours later, the cells were fixed with 1% paraformaldehyde and washed before being added to the coculture. For immune cell stimulation, splenocytes (5 × 105 cells/well) were incubated with prefixed target cells (B16-OVA, 5 × 104 cells/well) or one of the following peptides at 2 μg/mL: mgp100 (refs. 24–32 (link); AnaSpec), OVA (257–264; Invitrogen), TRP-2 (180–188; AnaSpec), or GFP (118–126; Biomatik). Twenty-four hours after incubation in a round-bottom 96-well plate, the concentration of IFNγ in the medium was assessed with a mouse IFNγ DuoSet ELISA kit (R&D Systems).
Jiang H., Shin D.H., Yi Y., Fan X., Gumin J., He J., Gillard A.G., Lang F.F., Gomez-Manzano C, & Fueyo J. (2023). Adjuvant Therapy with Oncolytic Adenovirus Delta-24-RGDOX After Intratumoral Adoptive T-cell Therapy Promotes Antigen Spread to Sustain Systemic Antitumor Immunity. Cancer Research Communications, 3(6), 1118-1131.
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6

Quantifying Tumor Cytokine Levels

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Tumor tissue was harvested and sonicated in the presence of radioimmunoprecipitation assay buffer (Sigma-Aldrich, R0278). After sonication, lysates were prepared by centrifugation, and protein concentration was then measured using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, 23227) according to the manufacturer’s protocol. IFN-γ and TNF-α levels in the tumor lysates were measured using a mouse IFN-γ DuoSet ELISA kit (R&D Systems, DY485) and a mouse TNF-α DuoSet ELISA kit (R&D Systems, DY410), respectively.
You G., Lee Y., Kang Y.W., Park H.W., Park K., Kim H., Kim Y.M., Kim S., Kim J.H., Moon D., Chung H., Son W., Jung U.J., Park E., Lee S., Son Y.G., Eom J., Won J., Park Y., Jung J, & Lee S.W. (2021). B7-H3×4-1BB bispecific antibody augments antitumor immunity by enhancing terminally differentiated CD8+ tumor-infiltrating lymphocytes. Science Advances, 7(3), eaax3160.
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7

Quantifying Bioactive Cytokine Binding

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The amount of bioactive IL12 bound to gold Nps was determined using an in vitro bioassay based on IL12-induced release of IFNγ from murine splenocytes [23 (link)]. IFNγ was detected using the mouse IFN-γ DuoSet ELISA kit (R&D System). The amount of bioactive TNF bound to gold Nps was determined using an in vitro bioassay based on TNF-induced cytolysis of murine L–M fibroblasts, as described previously [29 (link)].
Corti A., Sacchi A., Gasparri A.M., Monieri M., Anderluzzi G., Colombo B., Gori A., Mondino A, & Curnis F. (2021). Enhancement of doxorubicin anti-cancer activity by vascular targeting using IsoDGR/cytokine-coated nanogold. Journal of Nanobiotechnology, 19, 128.
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8

Measuring IFN-γ in Neuron-CD8+ T Cell Co-culture

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IFN-γ protein levels were measured from cryopreserved neuron-CD8+ T cell co-culture supernatant using the Mouse IFN-γ DuoSet ELISA kit from R&D Systems (R&D Systems, cat#DY48505, Minneapolis, MN) according to the manufacturer’s instructions.
Reagin K.L., Lee R.L., Cocciolone L, & Funk K.E. (2024). Antigen non-specific CD8+ T cells accelerate cognitive decline in aged mice following respiratory coronavirus infection. bioRxiv.
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9

Cytokine Profiling in Murine Serum

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Serum samples were collected as previously described. IFN-γ was detected in serum samples using the Mouse IFN-γ Duoset ELISA kit (R&D). Brain supernatants were inactivated by 1% FBS 1% Triton X-100 diluted in PBS and subjected to a 44-plex Luminex assay (Eve Technologies).
Balint E., Feng E., Giles E.C., Ritchie T.M., Qian A.S., Vahedi F., Montemarano A., Portillo A.L., Monteiro J.K., Trigatti B.L, & Ashkar A.A. (2024). Bystander activated CD8+ T cells mediate neuropathology during viral infection via antigen-independent cytotoxicity. Nature Communications, 15, 896.
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10

Quantifying IFN-γ in Tumor Lymph Nodes

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Tumor-draining lymph nodes were collected, homogenized for 30 s at 15 Hz using a TissueLyser II device (Qiagen, Hilden, Germany) and centrifuged at 12000×g for 5 min. The supernatant was collected to measure the amount of IFN-γ, using the Mouse IFN-γ DuoSet ELISA Kit (R&D Systems, Minneapolis, MN). The results were expressed as nmol/ml.
Salaroglio I.C., Gazzano E., Abdullrahman A., Mungo E., Castella B., Abd-elrahman G.E., Massaia M., Donadelli M., Rubinstein M., Riganti C, & Kopecka J. (2018). Increasing intratumor C/EBP-β LIP and nitric oxide levels overcome resistance to doxorubicin in triple negative breast cancer. Journal of Experimental & Clinical Cancer Research : CR, 37, 286.
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