Total bile acids test wako kit

The experiment was performed according to the methods of Huth et al. (2000) (link). LF (0, 5, and 10 mg/mL) or transferrin (10 mg/mL) was incubated with 0.5 mg/mL sodium taurocholate in 2 mL of phosphate buffer (pH 6.5) at 37°C for 2h with shaking. Unbound taurocholate was separated by ultrafiltration using a 10K MWCO membrane (Merck Millipore Corp., Darmstadt, Germany) and measured with the Total Bile Acids Test Wako kit (Wako Pure Chemical Industries, Ltd.).

Serum levels of total cholesterol and triglyceride were analyzed using the Cholesterol E-test Wako kit and Triglyceride E-test Wako kit, respectively (Wako Pure Chemical Industries, Ltd.).
Hepatic lipids were extracted with chloroform/methanol (2:1, v/v) according to the method of (Folch et al. 1957) . Extracted lipids were saponified in ethanolic KOH and nonsaponifiable lipids were extracted from the alkaline mixture with chloroform/methanol (2:1, v/v). Fecal steroids were extracted with hot ethanol. Quantification of hepatic cholesterol and fecal neutral D r a f t steroids was performed by gas chromatography (7890A instrument; Agilent Technologies, Santa Clara, CA, USA) using a HP-5 column (Agilent Technologies). 5α-Cholestane (Sigma-Aldrich) was used as an internal control. Fecal bile acids were analyzed using the Total Bile Acids Test Wako kit (Wako Pure Chemical Industries, Ltd.).