Ab213477

Urinary glucose and serum and urinary creatinine were measured using the colorimetric/fluorometric kits ab65333 (Abcam, Cambridge, UK) and K625 (Biovision, Milpitas, CA), respectively. Urinary cystatin C (ab201280; Abcam, Cambridge, UK), albumin (ab108792; Abcam, Cambridge, UK), and Kim-1 (ab213477; Abcam, Cambridge, UK) were measured by enzyme-linked immunosorbent assay (ELISA).

Blood was collected from the mice around PD30–PD40 (n = 4–10 per group) during the dissection procedure. The blood was left at RT for 30 minutes and centrifuged at 2,000 rcf. The resulting supernatant (serum) was transferred to a new 1.5 mL tube and stored at –20 °C. The serum samples were analyzed by IDEXX BioAnalytics. The evaluation was performed on a standard set of toxicity markers: glucose, total bilirubin, blood urea nitrogen, CK, creatinine, ALP, ALT, AST, GGT, globulin, albumin, and total protein. Urinary KIM-1 analysis was performed from urine collected from mice at PD30. The collected urine was centrifuged at 2,000 rcf at 4°C. The resulting supernatant was transferred to a new 1.5 mL tube and stored at –20°C until further use. Analysis was performed using a single-wash 90-minute sandwich ELISA protocol according to the manufacturer’s instructions (Abcam ab213477).