DuoSet ELISA kits (R&D Systems, Minneapolis, MN, and BD Biosciences, Oxford, Oxon, UK were used to assess serum cytokine levels according to the manufacturers’ instructions. Absorbance was read at 450 nm using a spectrophotometric ELISA plate reader (Anthos HTII; Anthos Labtec, Salzburg, Austria).
MILLIPLEX®MAP multi-analyte panels (Merck Millipore, Watford, Herts, UK) were used for simultaneous detection and quantification of eight biomarkers and cytokines/chemokine in rat urine, including NGAL, cystatin C, IL-18, MCP-1, clusterin, calbindin, osteopontin, and KIM-1. The same technique was used to determine serum levels of IL-18. Assays were performed according to the manufacturer’s protocols. The plate was read on a Bio-Plex 200 multiplex system (Bio-Rad, Hemel Hempstead, Herts, UK). Urine TIMP-2 and IGFBP7 were analysed by ELISA as described previously (6 (link)). Renal function (serum creatinine) was analyzed using the Jaffe assay by the Clinical Pathology laboratory at the Royal Free Hospital, London, UK. Where urine biomarkers were analyzed, matched creatinine values were used.
MILLIPLEX®MAP multi-analyte panels (Merck Millipore, Watford, Herts, UK) were used for simultaneous detection and quantification of eight biomarkers and cytokines/chemokine in rat urine, including NGAL, cystatin C, IL-18, MCP-1, clusterin, calbindin, osteopontin, and KIM-1. The same technique was used to determine serum levels of IL-18. Assays were performed according to the manufacturer’s protocols. The plate was read on a Bio-Plex 200 multiplex system (Bio-Rad, Hemel Hempstead, Herts, UK). Urine TIMP-2 and IGFBP7 were analysed by ELISA as described previously (6 (link)). Renal function (serum creatinine) was analyzed using the Jaffe assay by the Clinical Pathology laboratory at the Royal Free Hospital, London, UK. Where urine biomarkers were analyzed, matched creatinine values were used.