2.10. Evans blue dye leakage

UPARANT effects on vascular permeability were determined by quantifying Evans blue dye leakage extravasation. Evans blue dye was dissolved in normal saline (20 mg/mL) and filtered. Anaesthetized rats were injected with Evans blue dye through the femoral vein at 20 mg/kg. Sixty minutes later, each rat was perfused through the left cardiac ventricle with 15 mL of heparinized saline (4 U/mL) under constant peristaltic flow (10 mL/min) to purge out the circulating dye. Then, the kidneys were harvested, dissected and weighed. The Evans blue dye was extracted with formamide overnight at 65°C and read at 620 nm using a plate reader (Microplate Reader 680 XR; Bio‐Rad Laboratories).