DuoSet enzyme-linked immunosorbent assay (ELISA) kits (R&D Systems, Minneapolis, MN; BD Biosciences, Oxford, Oxon, United Kingdom) 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).
MILLIPLEXMAP multianalyte panels (Merck Millipore, Watford, Herts, United Kingdom) were used for simultaneous detection and quantification of eight biomarkers and cytokines/chemokine in rat urine, including neutrophil gelatinase-associated lipocalin (NGAL), cystatin C, interleukin (IL)-18, monocyte chemotactic protein (MCP)-1, clusterin, calbindin, osteopontin, and kidney injury molecule (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, United Kingdom). Urine tissue inhibitor of metalloproteinases-2 (TIMP-2) and insulin-like growth factor-binding protein 7 (IGFBP7) were analyzed 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, United Kingdom. Where urine biomarkers were analyzed, matched creatinine values were used.
MILLIPLEXMAP multianalyte panels (Merck Millipore, Watford, Herts, United Kingdom) were used for simultaneous detection and quantification of eight biomarkers and cytokines/chemokine in rat urine, including neutrophil gelatinase-associated lipocalin (NGAL), cystatin C, interleukin (IL)-18, monocyte chemotactic protein (MCP)-1, clusterin, calbindin, osteopontin, and kidney injury molecule (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, United Kingdom). Urine tissue inhibitor of metalloproteinases-2 (TIMP-2) and insulin-like growth factor-binding protein 7 (IGFBP7) were analyzed 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, United Kingdom. Where urine biomarkers were analyzed, matched creatinine values were used.
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