Mouse il 17 duoset elisa

The levels of serum-IgE were evaluated by using Mouse IgE Ready-Set-Go (eBioscience). The detection limit was 4 ng/ml and results were shown as ng/ml IgE. Concentration of IL-23 in LHG was measured with Mouse IL-23 DuoSet ELISA (R&D Systems). The limit of detection was 4 ng/ml and results were shown as ng/ml IL-23. Concentration of IL-17 in LHG was measured with Mouse IL-17 DuoSet ELISA (R&D Systems). The limit of detection was 4 ng/ml and results were shown as ng/ml IL-17.

Groups of three outbred 6-week-old female Swiss (CD-1) mice were intraperitoneally administered with 200 μg of either monoclonal anti-mouse IL-17A or mouse IgG1 isotype control antibodies one day prior to and 2 h before intranasal challenge with 9–47, as described above. Six hours of post-infection, lung tissue and bronchoalveolar lavage (BAL) were harvested. To quantify IL-17 levels, a mouse IL-17 DuoSet^® ELISA (R&D Systems) was performed on BAL samples in duplicate, according to the manufacturer’s instructions. Flow cytometry was performed on lung tissue samples, as described above, to quantify neutrophil levels after anti-mouse IL-17A treatment.

Cytokines in BAL were quantified using a cytometric bead array (CBA) kit mouse inflammation (BD Biosciences, San Diego, CA), according to the manufacturer's protocol. It was analyzed on a FACSCanto™ II (BD Biosciences, San Diego, CA) with FACSDiva software. IL-17 production was assessed according to the manufacturer's protocol (mouse IL-17 DuoSet ELISA, R&D Systems).