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MOESM3 of Cytokine production and phenotype of Histomonas meleagridis-specific T cells in the chicken

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posted on 2019-12-06, 04:45 authored by Julia Lagler, Taniya Mitra, Selma Schmidt, Alix Pierron, Eleni Vatzia, Maria Stadler, Sabine Hammer, Kerstin Mair, Beatrice Grafl, Patricia Wernsdorf, Fabienne Rauw, Bénédicte Lambrecht, Dieter Liebhart, Wilhelm Gerner
Additional file 3. Suitability of chicken IFN-γ-specific monoclonal antibodies specific for intracellular cytokine staining. A panel of six mAbs with either mouse IgG1 isotype (2B7, 11G5, 7E3, 12F12), or mouse IgG2a isotype (12F7) and mouse IgG2b isotype (12D4) was tested on PMA/ionomycin stimulated splenocytes. For each antibody, results are shown for the optimal quantity (clone 12F7: 150 ng, 2B7: 50 ng, 11G5: 12.5 ng, 7E3: 100 ng, 12D4: 250 ng, 12F12: 100 ng), initially identified in experiments with serial dilutions. Goat-anti-mouse isotype specific RPE-conjugated antibodies were applied afterwards for fluorescence labelling. Cells were pre-gated as described in Additional file 4A. Results are representative of four experiments with splenocytes from three different chickens.

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Veterinärmedizinische Universität Wien

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