IFNγ, IL-2 and granzyme B concentrations were measured in supernatants from in vitro activated human lymphocytes. BD OptEIA Set Human IFNγ (555142, BD Bioscience), human BD OptEIA Set Human IL-2 (555190, BD Bioscience) and Human Granzyme B ELISA development kit (3485-1H-6, Mabtech) were performed according to the manufacturer’s instructions.
Opteia set human ifn γ
Manufactured by BD
The BD OptEIA Set Human IFN-γ is a laboratory equipment product that is used for the quantitative measurement of human interferon-gamma (IFN-γ) in biological samples. It provides the necessary components to perform a sandwich enzyme-linked immunosorbent assay (ELISA) to detect and quantify IFN-γ levels.
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3 protocols using opteia set human ifn γ
1
Cytokine and Granzyme B Quantification
2
Cytokine Quantification in Cell Culture
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For the analysis of cytokine production in cell culture supernatants the following commercially available enzyme-linked immunosorbent assay (ELISA) kits have been used according to the manufacturers’ instructions: Human IL-22 DuoSet ELISA (DY782), Human TNFα DuoSet ELISA (DY210), Human IL-6 DuoSet ELISA (DY206), Human IL-10 DuoSet ELISA (DY217B), and Human CXCL10/IP-10 DuoSet ELISA (DY266) from R&D Systems (Minneapolis, MN) and BD OptEIA Set Human IFN-γ (555142) and BD OptEIA Set Human IL-4 (555194) from BD Biosciences. For the FLCN2 ELISA the stool was diluted in PBS at a concentration of 50 mg/mL, homogenized by vortexing for 5 minutes, and centrifuged for 10 minutes at 3000 g, and the supernatant was stored at –20°C until analyzed. FLCN2 was assayed using the Mouse Lipocalin-2/NGAL DuoSet ELISA (DY1857; R&D Systems). The samples were diluted as required and results calculated from the standard curve.
3
Cytokine Production Quantification
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Heparinized peripheral venous blood was diluted 1:10 with RPMI 1640 and incubated without stimulation or in the presence of either Phytohaemagglutinin (PHA, 10 µg/mL) or Streptococcal pyrogenic exotoxin A (SPEA, 250 ng/mL) at 37 °C and 5% CO2 for 3 and 5 days in sterile polypropylene tubes. Supernatants were obtained by centrifugation (10 min, 300× g) and stored at −80 °C. IFN-γ was quantified by ELISA using BD OptEIA Set Human IFN-γ (BD Biosciences, Heidelberg, Germany) according to the manufacturer’s instructions.
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