Profiler array rat xl cytokine array kit

After incubation of NR8383 cells with different silica particles (100 µg mL⁻¹) for 16 h, the supernatants were centrifuged at 300 g for 10 min and stored at -20 °C until the microarray analysis (Profiler Array Rat XL Cytokine Array Kit, Bio-Techne GmbH, Wiesbaden-Nordenstadt, Germany). The assay detected 79 different cytokines, growth factors and other mediators and enabled a semiquantitative analysis. The membrane-based sandwich immunoarray consists of a nitrocellulose membrane on which the captured antibodies are spotted as duplicated dots. Target proteins present in the sample bind to the capture antibodies and are detected with biotinylated detection antibodies and then visualized with chemiluminescent detection reagents. For analysis, the manufacturer's instructions were observed and chemiluminescence signals were detected and quantified by a microarray imager and the ImageQuantTL software (Amersham Imager 600 RGB, GE Healthcare Bio-Science, Uppsala, Sweden). For subsequent detailed analysis using Sandwich-ELISA Kits (R&D Systems Quantikine, Bio-Techne GmbH, Wiesbaden, Germany), 27 factors were selected based on the proteomic repertoire of NR8383 cells, as reported by Duhamel et al.^{58 (link)}.

After incubation of NR8383 cells with different ZnO particles (10 μg ZnO mL⁻¹) for 16 h, the supernatants were collected and centrifuged at 300g for 10 min and stored at − 20 °C until microarray analysis (Profiler Array Rat XL Cytokine Array Kit, Bio-Techne GmbH, Wiesbaden-Nordenstadt, Germany). The assay detected 79 different cytokines, growth factors, and other mediators and permitted a semi-quantitative analysis. The membrane-based sandwich immunoarray consisted of a nitrocellulose membrane on which the capture antibodies were spotted as duplicated dots. The target proteins in the sample were bound to the capture antibodies and detected with biotinylated detection antibodies, followed by visualization with chemiluminescent detection reagents. For analysis, the manufacturer’s instructions were observed and the chemiluminescence signals were detected and quantified by a microarray imager and the ImageQuantTL software (Amersham Imager 600 RGB, GE Healthcare Bio-Science, Uppsala, Schweden). For the subsequent detailed analysis, 27 factors were selected based on the proteomic repertoire of NR8383 cells (Duhamel et al. 2015 (link)).
Quantitative analyses were performed with cell culture supernatants after 16 h of exposure to ZnO particles (5 to 10 μg ZnO mL⁻¹) with Sandwich-ELISA Kits (R&D Systems Quantikine, Bio-Techne GmbH, Wiesbaden, Germany).