For the detection of soluble sIL-6R in sera of mice, the samples were diluted 1:15 in 1% BSA–PBS. The ELISA was performed as described67 (link). In brief, microtiter plates (Greiner Microlon) were coated with anti-murine IL-6-R pAB AF1830 (0.8 μg ml−1; R&D Systems) in PBS. After blocking, 100 μl aliquots of cell lysates or culture supernatants were added. IL-6R bound to the plate was detected by biotinylated goat anti-mouse IL-6-R pAB BAF1830 (working concentration, 0.2 μg ml−1; R&D Systems) followed by streptavidin–horseradish peroxidase (R&D Systems). The enzymatic reaction was performed with soluble peroxidase substrate (BM blue POD from Roche) at 37 °C and the absorbance was read at 450 nm on a SLT Rainbow plate reader (Tecan). For the detection of murine IL-6 in sera of mice, the samples were diluted 1:3 with 1% BSA–PBS.
Corresponding Organization : University of Veterinary Medicine Vienna
Other organizations :
Ludwig Boltzmann Institute for Cancer Research, University of Salzburg, Medical University of Vienna, Queen's University Belfast, Kiel University, New York University, University of Ulster, Austrian Academy of Sciences, CeMM Research Center for Molecular Medicine, University Medical Center of the Johannes Gutenberg University Mainz, Johannes Gutenberg University Mainz, University of Turin, Comprehensive Cancer Center Vienna, University of Tübingen, University of Cambridge, Swedish University of Agricultural Sciences, Innsbruck Medical University, Universität Innsbruck
Dilution of samples for murine IL-6 detection (1:3 in 1% BSA–PBS)
positive controls
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