Sigma bilirubin assay kit

Mice were fasted about 10 hours prior to blood collection. Blood was collected from the retro-orbital venous sinus using EDTA-coated blood-collecting capillaries. Mice were temporarily anesthetized using isoflurane before collecting blood. For nonterminal procedures about 200 µL was collected otherwise about 600 µL was collected. Blood from the capillary was transferred into BD microtainer tubes and then centrifuged according to the manufacturer’s protocol to separate the serum. Serum was then transferred into 1.5 mL microcentrifuge tubes and snap-frozen in liquid nitrogen before storing at −80 °C. Serum cholesterol, triglycerides, ALT, and AST activity were measured using colorimetric assay kits provided by Infinity (Thermo Scientific). Serum bilirubin, both direct and indirect, was measured using the Sigma Bilirubin Assay kit (MAK126 Sigma) according to the manufacturer’s protocol. Snap-frozen serum (100 µL) collected from control and acSRSF1 HKO mice were submitted to the Mouse Metabolic Phenotyping Core at the University of Cincinnati for serum analysis. A serum chemistry panel was obtained for each sample with measured concentrations of serum triglycerides, cholesterol, phospholipids, non-esterified fatty acids, glucose, ketones, ALT, and AST.