Mouse total ba assay kit

Full-length wide type recombinant human FGF19 (FGF19^WT) and the FGF1 variant (FGF1^ΔHBS) carrying triple mutations (Lys127Asp, Lys128Gln, and Lys133Val) were expressed in E.coli (BL21) and purified as described in the previous studies (Goetz et al., 2007 (link); Huang et al., 2017 (link)). Mouse Total BA Assay Kit was purchased from Crystal Chem INC. Alpha-naphthylisothiocyanate (ANIT) was purchased from Sigma Aldrich (St. Louis, MO, USA). Alanine transaminase (ALT) and aspartate transaminase (AST) Assay Kits were purchased from Sigma Aldrich (St. Louis, MO, USA) and performed in accordance with the protocols provided with manufacturers. All other reagents were of analytical grade.

Total BAs in the liver were quantified using the Mouse Total BA Assay Kit (Crystal Chem INC). Total BAs were extracted from the homogenate by agitating the tissue for two hours at 50 °C after it had been homogenized with 75% ethanol from individual livers. After being centrifuged, the extraction’s supernatants were collected and diluted in PBS for examination. The dilution parameters for each tissue extract were determined to ensure that the BA readings obtained using a mouse BA kit were within the linear range of the standard curve. The liver’s total number of BAs was added to estimate the size of the BA pool.