αifn γ

Manufactured by Thermo Fisher Scientific

Sourced in United States

αIFN-γ is a recombinant human interferon-gamma protein. It is a laboratory reagent commonly used in immunology research and development.

Automatically generated - may contain errors

Lab products found in correlation

7 protocols using αifn γ

Differentiation of Naive T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Naive T cells were isolated by sorting CD4⁺CD25⁻CD62L^highCD44^low cells from spleens and lymph nodes, differentiated under several Th cell conditions, and analyzed as described (Yang et al., 2008 (link)). The naïve T cells (5 × 10⁵ cells/well) were stimulated with the plate-bound α-CD3 (1 μg/ml) and the soluble α-CD28 (1 μg/ml). For Th0 differentiation, the cells were treated with 5 μg/ml α-IFN-γ (XMG1.2; BioXCell), 5 μg/ml α-IL-4 (11B11; BioXCell) and 30 U/ml IL-2. For Th1 differentiation, the cells were treated with 10 ng/ml mIL-12 (Peprotech), 5 μg/ml α-IL-4 and 30 U/ml IL-2. For Th2 differentiation, the cells were treated with 10 ng/ml mIL-4 (Peprotech), 5 μg/ml α-IFN-γ. For iTreg differentiation, the cells were treated with 1 ng/ml TGF-β1 (Peprotech), 5 μg/ml α-IFN-γ, 5 μg/ml α-IL-4 and 30 U/ml IL-2. For Th17 differentiation, the cells were treated with 0.5 ng/ml TGF-β1, 10 ng/ml IL-6 (Peprotech), 5 μg/ml α-IFN-γ, and 5 μg/ml α-IL-4. When indicated, 10 ng/ml IL-23 and 10 ng/ml IL-1β (Peprotech) were used for optimal Th17 cell differentiation.

Ichiyama K., Chen T., Wang X., Yan X., Kim B.S., Tanaka S., Ndiaye-Lobry D., Deng Y., Zou Y., Zheng P., Tian Q., Aifantis I., Wei L, & Dong C. (2015). The methylcytosine dioxygenase Tet2 promotes DNA demethylation and activation of cytokine gene expression in T cells. Immunity, 42(4), 613-626.

+ Open protocol

+ Expand

Differentiation of Naive T Cells into Th Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols

Naive T cells were isolated by sorting CD4⁺CD25⁻CD62L^highCD44^low cells from spleens and lymph nodes, differentiated under several Th cell conditions, and analyzed as described (Yang et al., 2008a (link)). Briefly, naïve T cells (5 × 10⁵ cells/well) were cultured at 37°C (5% CO₂) in complete medium. The cells were stimulated with the plate-bound α-CD3 (1 μg/ml; BioXCell) and the soluble α-CD28 (1 μg/ml; BioXCell). For Th0, the cells were treated with 30U/ml IL-2, 5 μg/ml α-IFN-γ (XMG1.2; BioXCell) and 5 μg/ml α-IL-4 (11B11; BioXCell). For Th1, the cells were treated with 30U/ml IL-2, 10 ng/ml IL-12 (Peprotech) and 5 μg/ml α-IL-4. For Th2, the cells were treated with 30U/ml IL-2, 10 ng/ml IL-4 (Peprotech) and 5 μg/ml α-IFN-γ. For Treg, the cells were treated with 30U/ml IL-2, 1 ng/ml TGF-β1 (Peprotech), 5 μg/ml α-IFN-γ and 5 μg/ml α-IL-4. For Th17, the cells were treated with 0.5 ng/ml TGF-β1, 10 ng/ml IL-1β (Peprotech), 10 ng/ml IL-6 (Peprotech), 10 ng/ml IL-23, 5 μg/ml α-IFN-γ and 5 μg/ml α-IL-4. For Th17(β), the cells were treated with 0.5 ng/ml TGF-β1, 10 ng/ml IL-6, 5 μg/ml α-IFN-γ and 5 μg/ml α-IL-4. When indicated, 10 μg/ml α-TGF-β (1D11; BioXCell) or ¹⁰ μM TGF-β receptor kinase inhibitor (SB431542; Calbiochem) were added. For Th17(23), the cells were treated with 10 ng/ml IL-1β, 10 ng/ml IL-6, 10 ng/ml IL-23, 5 μg/ml α-IFN-γ and 5 μg/ml α-IL-4.

Ichiyama K., Gonzalez-Martin A., Kim B.S., Jin H.Y., Jin W., Xu W., Sabouri-Ghomi M., Xu S., Zheng P., Xiao C, & Dong C. (2016). The microRNA-183-96-182 cluster promotes T helper 17 cell pathogenicity by negatively regulating transcription factor Foxo1 expression. Immunity, 44(6), 1284-1298.

+ Open protocol

+ Expand

In vitro Naive CD4+ T cell Polarization

Check if the same lab product or an alternative is used in the 5 most similar protocols

Naïve CD4⁺ T cells were sorted from the spleens and lymph nodes with the MACS CD4⁺CD62L⁺ T Cell Isolation (30–093-227) Kit (Miltenyi Biotec, Bergisch Gladbach, Germany). Cells were cultured under neutral (TH0) conditions in supplemented IMDM medium in the presence of activating antibodies (5 μg/ml plate-coated anti-CD3 and 1 μg/ml soluble anti-CD28) and iTreg polarizing cytokines: TGF-β [5 ng/ml], human IL-2 [20 ng/ml], αIL-4 [2 μg/ml], αIFN-γ [2 μg/ml] and αIL-12 [2 μg/ml].
Recombinant proteins (recombinant human IL-2 and TGF-β) and blocking antibodies (anti-mouse IL-4, anti-mouse IFN-γ, anti-mouse IL-12) for in vitro cell differentiation were purchased from eBioscience (San Diego, California,USA).

Thuille N., Siegmund K., Klepsch V., Schörgenhuber J., Danklmaier S., Leitges M, & Baier G. (2019). Loss-of-function phenotype of a PKCθT219A knockin mouse strain. Cell Communication and Signaling : CCS, 17, 141.

+ Open protocol

+ Expand

Detailed Workflow for Murine T Cell Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Isolated cells were stained with αCD4, αCD8, αCD3, αTCRβ, αCD44, αCD62L, αFoxP3, αIL-17 and αIFN-γ (eBioscience and Tombo). For FoxP3 and cytokine staining, cells were stained according to the manufacturer’s protocol form FoxP3 staining kit (Affimetrix/eBioscience). For cytokine staining, cells were stimulated with 50 ng/ml PMA and 1 μg/ml ionomycin in the presence of GolgiPlug (BD Bioscience) for 4 hours at 37°C and stained for surface markers followed by fixation/permeabilization and staining for cytokines. Stained cells were acquired with LSRII or LSR Fortessa cytometers; data was analyzed using FlowJo (Treestar) (BD Bioscience). For sorting of Treg cells, CD4+ T cells were enriched by using mouse CD4+ T cell isolation kit (Miltenyi) or Magnisort mouse CD4+ T cell enrichment kit (Affimetrix/eBioscience) and then cells were sorted by FACSAria (BD Bioscience).

Oh H., Grinberg-Bleyer Y., Liao W., Maloney D., Wang P., Wu Z., Wang J., Bhatt D.M., Heise N., Schmid R.M., Hayden M.S., Klein U., Rabadan R, & Ghosh S. (2017). An NF-κB transcription factor-dependent, lineage specific transcriptional program promotes regulatory T cell identity and function. Immunity, 47(3), 450-465.e5.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry of Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Flow-cytometric analysis of single-cell suspensions from spleen, lymph nodes (inguinal, axillary, cervical and mesenteric lymph nodes), thymus, whole blood, bone marrow, and TILs was performed using α-CD45, α-CD8, α-CD4, α-CD19, α-CD44, α-CD25, α-CD122, α-FoxP3, α-IFN-γ (all eBioscience), α-TNF-α, α-Ki67, α-CD3 (BioLegend), α-NK1.1 (BD Pharmingen). Stainings for intracellular FoxP3, Ki67, TNF-α, and IFN-γ were performed following the manufacturer’s instructions (eBioscience).

Jing H., Hettich M., Gaedicke S., Firat E., Bartholomä M, & Niedermann G. (2019). Combination treatment with hypofractionated radiotherapy plus IL-2/anti-IL-2 complexes and its theranostic evaluation. Journal for Immunotherapy of Cancer, 7, 55.

+ Open protocol

+ Expand

Monoclonal Antibody-Induced Cytokine Inhibition

Check if the same lab product or an alternative is used in the 5 most similar protocols

Monoclonal antibodies were applied to the vaccinated group to induce the functional inhibition of IFN-γ or IL-17A. Purified monoclonal anti-mouse IFN-γ (αIFN-γ; eBioscience; 16-7311-85) or anti-mouse IL-17A (αIL-17A; eBioscience; 16-7173-85) was prepared using endotoxin-free PBS. Two days before infection, vaccinated mice were injected with 100 μg (i.p.; 100 μL) of αIFN-γ, αIL-17A, or both. The vaccinated and nonvaccinated control groups were injected with the nonspecific isotype control IgG [100 or 200 μg (i.p.)] at doses comparable to those administered to the single inhibition group and the combined treatment group.

Yang L., Cai C., Feng Q., Shi Y., Zuo Q., Yang H., Jing H., Wei C., Zhuang Y., Zou Q, & Zeng H. (2016). Protective efficacy of the chimeric Staphylococcus aureus vaccine candidate IC in sepsis and pneumonia models. Scientific Reports, 6, 20929.

+ Open protocol

+ Expand

Differentiation and Transduction of γδT17 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Pooled spleen and LN cells were cultured at 1 × 10⁶ cells per ml in RPMI 1640 containing 10% FCS, antibiotics, 1 × Glutamax (Gibco), 10 mM HEPES (SA Pathology), 1 mM sodium pyruvate, 54 pM β-mercaptoethanol and 1 × non-essential amino acids (Gibco) with 5 ng ml⁻¹ recombinant (r)IL-23 (eBioscience), 5 ng ml⁻¹ rIL-1β (Miltenyi Biotec) and 10 μg ml⁻¹ α-IFN-γ (BioXCell) in 96-well round-bottom plates coated with 1 μg ml⁻¹ α-TCR-γδ (clone GL3; Biolegend) for 3 days. Cells were washed and re-seeded on fresh plastic at 1 × 10⁶ cells per ml for a further 3 days as above without TCR-γδ stimulation. Cells were then washed and re-seeded in 20 ng ml⁻¹ rIL-7 (Peprotech) and 10 μg ml⁻¹ α-IFN-γ for a further 3 days. pMIG, pMIG-Rorc and pMIG-Ccr6 (cloned from mouse Ccr6 cDNA) were transfected into EcoPack 2 293 cells (Clontech; mycoplasma free) with Lipofectamine 2000 (ThermoFisher), and supernatant collected after 48 h. γδT17 cells at days 4 and 5 of culture were centrifuged at 2,500 r.p.m. (30 °C for 1.5 h) in supernatant with 8 μg ml⁻¹ polybrene (Sigma) in flat-bottom 96 well trays before being returned to culture.

McKenzie D.R., Kara E.E., Bastow C.R., Tyllis T.S., Fenix K.A., Gregor C.E., Wilson J.J., Babb R., Paton J.C., Kallies A., Nutt S.L., Brüstle A., Mack M., Comerford I, & McColl S.R. (2017). IL-17-producing γδ T cells switch migratory patterns between resting and activated states. Nature Communications, 8, 15632.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!