Anti ifn γ apc clone b27

Manufactured by BD

The Anti-IFN-γ APC (clone: B27) is a monoclonal antibody that binds to the interferon-gamma (IFN-γ) protein. This antibody is conjugated with allophycocyanin (APC), which is a fluorescent dye. The primary function of this product is to detect and quantify IFN-γ in various experimental settings.

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Lab products found in correlation

Brefeldin a, by Merck Group (1 mentions) Facscanto 2, by BD (1 mentions) Sp600125, by Abcam (1 mentions) Anti il 13 pe clone jes10 5a2, by BD (1 mentions) Pom 1, by Bio-Techne (1 mentions) Anti ido 1apc, by R&D Systems (1 mentions) Cflow plus, by BD (1 mentions) Anti granzyme b pe clone gb11, by BD (1 mentions) Accuri c6, by BD (1 mentions) Anti cd8 fitc clone rpat8, by BD (1 mentions) Zombie red viability dye, by BioLegend (1 mentions) Brefeldin a, by BioLegend (1 mentions) Ionomycin, by Merck Group (1 mentions) Ultra low attachment 96 well plate, by Corning (1 mentions)

Close spelling variants of this product

IFN-γ APC (B27, IFN-γ (clone B27, IFN-γ (B27)( Anti-IFN-γ-APC IFN-γ-APC Anti-IFN-γ Clone B27 IFN-γ

4 protocols using anti ifn γ apc clone b27

NK Cell Cytotoxicity Assay

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Freshly isolated PBMCs and their in vitro cultured counterparts were stimulated with Raji cells (2:1), opsonized or not with the minimum saturating dose of rituximab (1 μg/1 × 10⁶) or obinutuzumab (0.1 μg/1 × 10⁶), or with K562 targets, for 6 h at 37°C in the presence of PE-conjugated anti-CD107a mAb (clone: H4A3, cat #: 555801; BD Biosciences) and 50 µM Monensin (Golgi-stop; cat #: M5273; Merck). After the first hour, 10 µg/ml Brefeldin A (cat #: B7651; Merck) was added. At the end of stimulation, cells were washed with 5 mM EDTA containing PBS and then stained as earlier, adding the anti-IFN-γ APC (clone: B27, cat #: 554702; BD Biosciences) after permeabilization.
Samples were analyzed with a FACSCanto II (BD), and data were obtained with FlowJo vX.0.7 (TreeStar) software. Where required, twin samples were added and stained with isotype control mAb, used to set the threshold for antigen positivity. The values of cell counts and cytofluorimetric analysis percentages were used to obtain the absolute number of different NK cell subsets.

Capuano C., Battella S., Pighi C., Franchitti L., Turriziani O., Morrone S., Santoni A., Galandrini R, & Palmieri G. (2018). Tumor-Targeting Anti-CD20 Antibodies Mediate In Vitro Expansion of Memory Natural Killer Cells: Impact of CD16 Affinity Ligation Conditions and In Vivo Priming. Frontiers in Immunology, 9, 1031.

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Multiparameter Analysis of DC Activation

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We used: PE-labeled antibodies to CD40 (clone 5C3), CD80 (clone L307.4), CD83 (clone HB15e), CD39 (clone TU66), PDL-1 (clone MIH1), as well as anti-CD4-APC (clone SK3) all from Becton Dickinson: (BD), anti-CD86-PE (clone HA5.2B7, Beckman Coulter), anti-IL-12p35/p70-PE (clone REA121, Miltenyi Biotech), anti-IL-12/IL-23p40-APC (clone C11.5, BioLegend), anti-IL-10-PE (clone JES3-19F1, BD), anti-IFN-γ-APC (clone B27, BD), anti-IL-13-PE (clone JES10-5A2, BD), anti-IDO-1-APC (clone 70083, R&D), anti-HO-1 (HO-1-1, Thermoscientific). For blocking experiments, we used rat neutralizing Ab to IL-10 (10 μg/ml, BD), IL-10R (10 μg/ml, R&D Systems), TLR2, TLR6, and Rat IgG control (20 μg/ml, InvivoGen), the JNK inhibitor SP600125 (10 μM, Abcam), the inhibitor of the CD39 ecto-nucleotidase activity Pom1 (10 μM, Tocris) and the IDO1 inhibitor (400 μM, Sigma Aldrich). DC were incubated with the inhibitor for 45 min before bacteria addition.

Alameddine J., Godefroy E., Papargyris L., Sarrabayrouse G., Tabiasco J., Bridonneau C., Yazdanbakhsh K., Sokol H., Altare F, & Jotereau F. (2019). Faecalibacterium prausnitzii Skews Human DC to Prime IL10-Producing T Cells Through TLR2/6/JNK Signaling and IL-10, IL-27, CD39, and IDO-1 Induction. Frontiers in Immunology, 10, 143.

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Flow Cytometry Analysis of Lymphocyte Markers

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All flow cytometry experiments were run on an Accuri C6 (BD Biosciences, Inc.) flow cytometer. Antibodies used were as follows: anti-BR3 PE, clone 11C1, BD Biosciences; anti-TACI APC, clone 165004, and anti-BCMA APC, goat pAb FAB193A, both from RnD Systems; anti-CD25 FITC-Violet and APC, clone 3H3, Miltenyi Biotec; anti-CD69 FITC, clone FN50, eBioscience; anti-IFN-γ APC, clone B27, BD Biosciences; anti-Granzyme B PE, clone GB11, BD Biosciences; anti-CRTAM PE, clone Cr24.1, Biolegend, Inc; anti-BAFF APC, clone 1D6, BD Biosciences; anti-BCMA APC, pAb FAB193A, RnD Systems, Inc; anti-cleaved PARP (Asp214) PE, clone XF21-852, BD Biosciences; anti-active Caspase 3 FITC, clone C92-605, BD Biosciences. For intracellular staining of IFN-γ, cells stimulated with plate-bound anti-CD3/CD28 for 12 hours were cultured in the presence of Brefeldin A for another 4-6 hours. Cells were then fixed and permeabilized using reagents from RnD Systems (FC004 Fixation Buffer, FC005 Permeabilization Buffer) according to manufacturer’s instructions. Analyses were performed using CFlowPlus (BD Biosciences) and/or FlowJo (TreeStar, Inc.) software.

Bloom D.D., Reshetylo S., Nytes C., Goodsett C.T, & Hematti P. (2018). Blockade of BAFF Receptor BR3 on T cells Enhances Their Activation and Cytotoxicity. Journal of immunotherapy (Hagerstown, Md. : 1997), 41(5), 213-223.

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Cytokine Profiling of Activated PBMCs

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PBMCs were plated at 1–3×10⁶ cells per well in ultra-low-attachment 96-well plates (Corning) in IMDM supplemented with brefeldin A (420601; BioLegend), 50 ng/mL PMA, and 0.5 μg/mL ionomycin (Sigma) and incubated at 37°C for 4 hours. The cells were washed, stained with Zombie Red viability dye (423109; BioLegend), and surface stained with anti–CD8-FITC (clone RPA-T8; BD). The cells were fixed with 4% PFA and permeabilized using 0.3% Triton X-100 in PBS and thereafter stained with anti–IFNγ-APC (clone B27; BD) and anti–IL-2-BV421 (clone MQ1-17H12; BioLegend).

Virtakoivu R., Rannikko J.H., Viitala M., Vaura F., Takeda A., Lönnberg T., Koivunen J., Jaakkola P., Pasanen A., Shetty S., de Jonge M.J., Robbrecht D., Ma Y.T., Skyttä T., Minchom A., Jalkanen S., Karvonen M.K., Mandelin J., Bono P, & Hollmén M. (2021). Systemic Blockade of Clever-1 Elicits Lymphocyte Activation Alongside Checkpoint Molecule Downregulation in Patients with Solid Tumors: Results from a Phase I/II Clinical Trial. Clinical Cancer Research, 27(15), 4205-4220.

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