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Anti mouse ifn γ apc

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The Anti-mouse IFN-γ APC is a fluorescent-labeled antibody that binds to mouse interferon-gamma (IFN-γ). This product is designed for use in flow cytometry applications to detect and quantify IFN-γ-producing cells.

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Lab products found in correlation

Anti mouse cd8α pe, by Thermo Fisher Scientific (3 mentions) Anti human ifn γ pe, by Thermo Fisher Scientific (2 mentions) Anti mouse cd3 percp eflour710, by Thermo Fisher Scientific (2 mentions) Permeabilization buffer, by Thermo Fisher Scientific (2 mentions) Anti mouse cd25 apc, by Thermo Fisher Scientific (2 mentions) Facscalibur, by BD (1 mentions) Anti mouse cd11c apc, by Thermo Fisher Scientific (1 mentions) Mouse igg1 κ isotype control, by Thermo Fisher Scientific (1 mentions) Easysep mouse cd8 t cell isolation kit, by STEMCELL (1 mentions) Anti mouse cd19 apc, by Thermo Fisher Scientific (1 mentions) Rat igg1 κ isotype control ebrg1, by Thermo Fisher Scientific (1 mentions) Easysep mouse nk cell isolation kit, by STEMCELL (1 mentions) Fixation permeabilization concentrate and diluent, by Thermo Fisher Scientific (1 mentions) Pe conjugated anti mouse rat foxp3, by Thermo Fisher Scientific (1 mentions) Facscanto 2 flow cytometer, by BD (1 mentions) Gentlemacs dissociator, by Miltenyi Biotec (1 mentions) Golgiplug, by BD (1 mentions) Ic fixation buffer, by Thermo Fisher Scientific (1 mentions) Anti mouse cd3 percp efluor 710, by Thermo Fisher Scientific (1 mentions) Il 17 pe, by Thermo Fisher Scientific (1 mentions)

9 protocols using anti mouse ifn γ apc

1

IFN-gamma Release Assay for CD8+ T Cells

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IFN-γ release of induced CD8+ T cells was measured by the intracellular cytokine staining assay according to our former study [31 (link), 34 (link)]. Briefly, the induced CTLs (1 × 106) and T2A2 cells loaded with/without corresponding peptide (1 × 106) were incubated by adding protein transport inhibitor (containing brefeldin A) for 5 hours at 37°C. Afterward, cells were harvested, transferred to 1.5 mL EP tubes, and washed by PBS (contain 2% FBS). Cells were then stained for cell surface markers (anti-human CD3 PerCP-eFlour (Clone: OKT3, 46-0037-42, eBioscience, USA)/anti-mouse CD3 PerCP-eFlour710 (Clone: 17A2, 46-0032-80, eBioscience, USA) and anti-human CD8α APC (Clone: SK1, 17-0087-42, eBioscience, USA)/anti-mouse CD8α PE (Clone: 53-6.7, 12-0081-82, eBioscience, USA)) for 30 min at 4°C. Afterward, the cells were fixed by fixation buffer for another 30 min at room temperature and washed twice with permeabilization wash buffer. Anti-human IFN-γ PE (Clone: 4S.B3, 12-7319-42, eBioscience, USA) or anti-mouse IFN-γ APC (Clone: XMG1.2, 17-7311-81, eBioscience, USA) was added to those tubes for intracellular staining for 30 min at 4°C. After being washed twice with permeabilization wash buffer, the IFN-γ release of CD8+ T cells was analyzed by flow cytometry (FACS Calibur, BD Bioscience, USA).
Lin Y., Dong Y., Gao Y., Shi R., Li Y., Zhou X., Liu W., Li G., Qi Y, & Wu Y. (2021). Identification of CTL Epitopes on Efflux Pumps of the ATP-Binding Cassette and the Major Facilitator Superfamily of Mycobacterium tuberculosis. Journal of Immunology Research, 2021, 8899674.
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2

Comprehensive Immune Cell Profiling

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Anti-human CD45 FITC (HI30), anti-human TIGIT APC (MBSA43), anti-human PD-1 PE (MIH4), anti-mouse TIGIT PE (GIGD7), anti-mouse PVR APC (TX56), anti-mouse PD-1 PE (J43), anti-mouse PD-L1 PE (MIH5), anti-mouse CD45 FITC (30-F11), anti-mouse CD3 PerCP-eFluor710 (17A2), anti-mouse CD8α PE (53-6.7) anti-mouse CD49b PE/APC (DX5), anti-mouse CD19 APC (eBio1D3), anti-mouse CD11c APC (N418), anti-mouse CD11b APC (M1/70), anti-mouse Ly-6G(Gr-1) PE- Cyanine7 (RB6-8C5), anti-mouse F4/80 PerCP-Cyanine5.5 (BM8), anti-mouse IFN-γ APC (XMG1.2), mouse IgG1κ isotype control (P3.6.2.8.1), rat IgM isotype control (eBR2M), rat IgG2α κ isotype control (eBR2a) and rat IgG1 κ isotype control (eBRG1) antibodies were purchased from eBioscience. Anti-mouse TIGIT APC (1G9) was purchased from BioLegend. Antibodies anti-asialo-GM1 (catalog 986-10001) (Wako Chemicals GmbH, Germany) and rabbit IgG control (I8140) (Sigma) were used for NK cell depletion. EasySep mouse CD8+ T cell isolation kit (catalog 19853) and EasySep mouse NK cell isolation kit (catalog 19855) (STEMCELL) were used for cell sorting.
Zhou X.M., Li W.Q., Wu Y.H., Han L., Cao X.G., Yang X.M., Wang H.F., Zhao W.S., Zhai W.J., Qi Y.M, & Gao Y.F. (2018). Intrinsic Expression of Immune Checkpoint Molecule TIGIT Could Help Tumor Growth in vivo by Suppressing the Function of NK and CD8+ T Cells. Frontiers in Immunology, 9, 2821.
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3

Epirubicin Effects on Tumor-Infiltrating Lymphocytes

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At day 0, female BALB/c mice (8 mice per group) were inoculated subcutaneously with CMS5a cells into the right inguinal region. Epirubicin (0.1, 0.3 or 1 mg/kg) or saline was given at days 3, 5 and 7 intravenously. On day 8, mice were euthanized and tumors were removed. Tumor-infiltrating lymphocytes (TIL) were dissociated from tumors using a gentleMACS dissociator according to the manufacturer’s instructions (Miltenyi Biotec). The collected cells were seeded in 24-well plates and stimulated with phorbol 12-myristate 13-acetate (PMA) and Ionomycin at 37°C for 1 h, and then cultured for 6 h with GolgiPlug™ (BD Biosciences). The cells were harvested and stained with PreCP-Cy™5.5 rat anti-mouse CD4 antibody (BD Pharmingen) and V500 rat anti-mouse CD8a antibody (BD Horizon) at 4°C for 15 min. The stained cells were fixed with Fixation/Permeabilization Concentrate and Diluent (1:3, eBioscience) at 4°C overnight. After washing, Permeabilization Buffer (eBioscience) was added and the fixed cells were stained with PE-conjugated anti-mouse/rat Foxp3 (eBioscience), anti-mouse IFN-γ-APC (eBioscience), and PE-conjugated anti-mouse IL-2 (BioLegend) antibodies. The stained cells were analyzed using a FACS Canto II flow cytometer (Becton Dickinson).
Kashima H., Momose F., Umehara H., Miyoshi N., Ogo N., Muraoka D., Shiku H., Harada N, & Asai A. (2016). Epirubicin, Identified Using a Novel Luciferase Reporter Assay for Foxp3 Inhibitors, Inhibits Regulatory T Cell Activity. PLoS ONE, 11(6), e0156643.
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4

Evaluating CTL Response to MTA1 Stimulation

Cited 1 time
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To test the protein MTA1(1–283) stimulation of CTL response in vivo, IFN-γ release of induced CTLs was determined by the intracellular cytokine staining assay according to our former study [32 (link)]. In brief, on day 14, spleen lymphocytes were harvested and stimulated by peptide pool (10 μg/ml) in vitro for another 6 days. Then, restimulated splenocytes were incubated with peptide-loaded T2 cells or T2 cells only for 3 h. Subsequently, protein transport inhibitor cocktail was added to splenocytes for another 4 h. Then, those cells were stained with Anti-mouse CD3 PerCP-eFluor® 710 (clone: 17A2, eBioscience) and Anti-mouse CD8α PE (clone: 53–6.7, eBioscience) at 4°C for 30 min. And those cells were fixed by IC fixation buffer (cat no: 00-8222, eBioscience) at room temperature for another 30 min; then those cells were washed twice by permeabilization buffer (cat no: 00-8333-56, eBioscience). After, those cells were incubated with Anti-mouse IFN-γ APC (clone: XMG1.2, eBioscience) at 4°C for 30 min. Those cells were washed twice by permeabilization buffer and analyzed by flow cytometry.
Wu Y., Zhai W., Zhou X., Wang Z., Lin Y., Ran L., Qi Y, & Gao Y. (2018). HLA-A2-Restricted Epitopes Identified from MTA1 Could Elicit Antigen-Specific Cytotoxic T Lymphocyte Response. Journal of Immunology Research, 2018, 2942679.
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5

Macrophage-Derived Soluble Factors Modulate T Cell Activation

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The RAW 264.7 cells with a density of 2*105/well were cultured in 6-well plates, and after cell adhesion the medium was replaced with CS ionic solution in different dilutions (1/8, 1/16, 1/32, 1/64, and 1/128). The cell culture supernatant was collected after 72 h, and labeled as CM-control, CM-1/8CS, CM-1/6CS, CM-1/32CS, CM-1/64CS and CM-1/128CS. The complete medium was used as a control.
The naïve T cells were sorted from mouse monocytes by using mouse CD4 (L3T4) magnetic bead antibody, which was close to 100% purity (Fig. S2). CD28 mouse antibody (BioCell) was added into 6-well plates and incubated for 2 h. Then, the solution was removed and naïve T cells were added into the well, followed by the addition of CS ionic solution with different dilutions (1/8, 1/16, 1/32, 1/64, and 1/128), or supernatant of macrophage cultured with CS ionic solutions (CM-control, CM-1/8CS, CM-1/16CS, CM1/32CS, CM-1/64CS, CM-1/128CS).
Meanwhile, naïve T cells were also activated by adding CD3 mouse antibody (BioCell), and collected after 48 h stimulation. Then, surface staining with anti-mouse CD4-FITC was performed. After fixation and membrane rupture, cells were intracellularly stained with anti-mouse IFNγ-APC, IL17-PE, or mouse FOX-P3-APC (eBioscience, USA), and the stained cells were analyzed by flow cytometry.
Chen T., Zhang Z., Weng D., Lu L., Wang X., Xing M., Qiu H., Zhao M., Shen L., Zhou Y., Chang J, & Li H.P. (2021). Ion therapy of pulmonary fibrosis by inhalation of ionic solution derived from silicate bioceramics. Bioactive Materials, 6(10), 3194-3206.
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6

Comprehensive Immunophenotyping Assays

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For the T2A2 cell-binding assay, monoclonal antibody HLA-A2 PE-Cyanine7 (clone: BB7.2, # 343314) was purchased from BioLegend (San Diego, CA, USA). For intracellular cytokine staining assay, monoclonal antibody anti-human CD3 eflour450 (clone: OKT3, #48-0037), anti-human CD8α PerCP-eFlour 710 (clone: SK1, #46-0087), anti-human IFN-γ PE (clone: 4S.B3, #12-7319), anti-human granzyme B PE (clone: GB11, #12-8899), anti-mouse CD3 PerCP-eFlour 710 (clone: 17A2, #46-0032), anti-mouse CD8α PE (clone: 53-6.7, #12-0081), anti-mouse IFN-γ APC (clone: XMG1.2, #17-7311), and anti-mouse granzyme B eFlour 450 (clone: NGZB, #48-8898) were purchased from eBioscience (San Diego, CA, USA). To detect the mature cells (DCs), anti-human CD80 PerCP-eFlour 710 (clone: 2D10.4, #46-0809), anti-human CD86 PE (clone: IT2.2, #12-0869), and anti-human HLA-DR APC (clone: LN3, #17-9956) were purchased from eBioscience (USA).
Wang Z., Ran L., Chen C., Shi R., Dong Y., Li Y., Zhou X., Qi Y., Zhu P., Gao Y, & Wu Y. (2021). Identification of HLA-A2-Restricted Mutant Epitopes from Neoantigens of Esophageal Squamous Cell Carcinoma. Vaccines, 9(10), 1118.
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7

Multiparameter Flow Cytometry Panel

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Streptavidin-PE (SA-PE), anti-mouse-CD45-FITC (30-F11), anti-mouse CD11c-PerCP-efluor 450 (N418), anti-mouse CD11b-APC (M1/70), anti-mouse-CD3-PerCP-eFlour710 (17A2), anti-mouse-CD8-APC (53-6.7), Rat IgG2a-APC kappa Isotype Control (eBR2a), anti-mouse-CD3-FITC (17A2), anti-mouse-CD8-PE (53-6.7), anti-mouse-IFN-γ-APC (XMG1.2), anti-mouse-Perforin-APC (eBioMAK-D), anti-mouse-Granzyme B-PerCP-eFlour710 (NGZB), Rat IgG1-APC kappa Isotype Control (eBRG1), Rat IgG2a-PerCP-eFlour710 kappa Isotype Control (eBR2a), anti-mouse-OVA257-264 (SIINFEKL/H-2Kb)-PE (eBio25-D1.16), Mouse IgG1-PE kappa Isotype Control (P3.6.2.8.1), anti-mouse-IL-21-PE (FFA21), Rat IgG2a-PE kappa Isotype Control (eBR2a), anti-mouse-Clec9a-PE (42D2), anti-mouse CD11c-APC (N418) and Rat IgG1-PE kappa Isotype Control (eBRG1) were purchased from eBioscience (San Diego, CA, USA). The anti-mouse/rat XCR1-Brilliant Violet 650TM (ZET) and anti-mouse/rat XCR1-APC (ZET) were purchased from Biolegend (San Diego, CA, USA). The anti-mouse IL-21 (FFA21) neutralized antibody was purchased from Thermo Fisher Scientific (Waltham, MA, USA). The phospho-Syk (Tyr323) antibody was purchased from CST (Boston, USA). The Anti-Histone H3 antibody was purchased from Abcam (Cambridge, United Kingdom).
Gou S., Wang S., Liu W., Chen G., Zhang D., Du J., Yan Z., Wang H., Zhai W., Sui X., Wu Y., Qi Y, & Gao Y. (2021). Adjuvant-free peptide vaccine targeting Clec9a on dendritic cells can induce robust antitumor immune response through Syk/IL-21 axis. Theranostics, 11(15), 7308-7321.
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8

Mouse MOG35-55 Peptide-Induced Neuroinflammation

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Rat MOG35–55 peptides were purchased from Biosynth International (Naperville, IL, USA) and the purity of the peptide was >95%. The sequence of MOG35–55 was MEVGWYRSPFSRVVHLYRNGK. FITC anti-mouse CD3, PE/Cy7 anti-mouse CD4, APC/Cy7 anti-mouse CD44, PerCP/Cy5.5 anti-mouse CD11b antibodies, and LEAF Purified anti-mouse IL-6 were purchased from BioLegend (San Diego, CA,USA). Alexa Fluor700 anti-mouse CD45.2, PE anti-mouse CD69, APC anti-mouse CD25, PE anti-mouse IL-17A, and APC anti-mouse IFN-γ antibodies were purchased from eBioscience (San Diego, CA, USA). PE/Cy5 anti-mouse CD8α antibodies were purchased from BD Pharmingen (Basel, Switzerland). Goat anti-mouse ionized calcium-binding adaptor molecule-1 (IBA1) antibody was obtained from abcam (Cambridge, UK). Rat anti-mouse CD45, Rabbit anti-mouse CCL-2 antibody, and CCL-2 (rat recombinant) were purchased from AbD Serotec (Raleigh, NC, USA). Rabbit anti-mouse CCR2 antibody was purchased from Abcam (Cambridge, UK). 2-((1-benzyl-indazol-3-yl) methoxy)-2-methyl propionic acid (Bindarit) was synthesized by and obtained from Angelini (Angelini Research Center-ACRAF, Italy).
Ji Z., Wu S., Xu Y., Qi J., Su X, & Shen L. (2019). Obesity Promotes EAE Through IL-6 and CCL-2-Mediated T Cells Infiltration. Frontiers in Immunology, 10, 1881.
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9

Th Cell Subset Profiling Protocol

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The spleen and MLNs were aseptically separated, and single-cell suspensions were prepared as described previously [14 (link)–16 (link)]. Single-cell suspensions from the spleen and MLNs were stimulated with a brefeldin/monensin mixture [Brefeldin A; Multisciences (Lianke) Biotech, Hangzhou, China] and a phorbol-12-myristate-13-acetate/ionomycin mixture [Multisciences (Lianke) Biotech] for 5 h. Fluorescein isothiocyanate (FITC)-anti-mouse-CD4 (eBioscience, Thermo Fisher Scientific, Waltham, MA, USA) was used for surface staining. APC-anti-mouse-IFN-γ (eBioscience, Thermo Fisher Scientific), APC-anti-mouse-IL-4 (BioLegend, San Diego, CA, USA) and PE-anti-mouse-IL-17A (Biolegend) were utilized for intracellular staining after the membrane was ruptured by fixation/permeabilization (BD Biosciences, Franklin Lakes, NJ, USA) to analyze Th1, Th2 and Th17 cells as described previously [16 (link), 19 (link)]. Mononuclear cells were stained with FITC-anti-mouse-CD4, APC-anti-mouse-CD25 and PE-anti-mouse Foxp3 by using a mouse regulatory T cell staining kit (eBioscience) to evaluate regulatory T (Treg) cells. All samples were analyzed using a BD FACSCcanto flow cytometer (BD Biosciences), and data were analyzed with Flowjo v10.0.7 software (Tree Star, Ashland, OR, USA).
Shan W., Zhang W., Xue F., Ma Y., Dong L., Wang T., Zheng Y., Feng D., Chang M., Yuan G, & Wang X. (2021). Schistosoma japonicum peptide SJMHE1 inhibits acute and chronic colitis induced by dextran sulfate sodium in mice. Parasites & Vectors, 14, 455.
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