Il 6 duoset kit

Quantification of IL-6 production was determined using a commercially available ELISA (IL-6 DuoSet kit, R&D Systems) following the manufacturer’s instructions. Cell culture supernatants were diluted 1:200 with assay buffer before being read on a BioTek EL808 microtiter plate reader (BioTek, Swindon, UK).

Quantification of IL-6 production was determined using a commercially available ELISA (IL-6 DuoSet kit, R&D Systems, DY206) following the manufacturer’s instructions. Cell supernatants were diluted 1:200 with assay buffer before being read on a BioTek EL808 microtiter plate reader (BioTek, Swindon, UK).

IL-6 serum concentrations were determined using the mouse IL-6 DuoSet Kit (DY406, R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instruction. In brief: Microplates were coated with 2 µg/mL Capture Antibody in PBS O/N at RT. After blocking with 1% BSA in PBS standards, blanks and samples were added, 100 µL, respectively. Murine serum samples were used undiluted. After incubation (O/N, 4 °C) biotinylated Detection Antibody (75 ng/mL) and subsequently Strepatavidin–HRP in blocking buffer was added. Wells were incubated with TMB substrate (555214, BD Biosciences, San Diego, CA, USA) and color development was stopped by adding 1 M H₂SO₄. The Absorbance at 450 nm was measured using a microplate reader (ThermoScientific, Waltham, MA, USA).