Bcg bromocresol green albumin assay kit

Mice were euthanized (500 mg/kg Ketamine + 50 mg/kg Xylazine) and BALF was collected through catheter insertion intratracheally, injecting and retracting 1 ml of ice-cold PBS. BALF was centrifuged at 300 × g for 5 min at 4°C and supernatants were stored at −80°C. Pelleted cells in the BALF were counted and analysed by flow cytometry to determine the frequency of Ter119⁺ RBCs on day 6 post infection unless otherwise indicated. Frequencies of immune cell populations were quantified by flow cytometry. Concentrations of IFN-α/β were measured using 2-Plex ProcartaPlex (Invitrogen). IFN-λ was measured by ELISA (R&D). Total protein concentrations in BALF were determined using the Pierce BCA Protein Assay Kit (Thermo Scientific) according to manufacturer’s instructions. Serum albumin was quantified using a BCG (Bromocresol Green) Albumin Assay Kit (Sigma) according to manufacturer’s instructions. The ProcartaPlex Cytokine and Chemokine Mouse 36-Plex (eBioscience) was used to assess cytokine concentrations in BALF read on a Luminex 100 (BioRad).

Fasting plasma insulin levels were measured in samples from 6 hour fasted mice by using an insulin ELISA kit (Mercodia, Uppsala, Sweden) following manufacturers’ protocol. Plasma leptin and adiponectin levels of the DIO mice were measured using a mouse leptin ELISA Kit (ChrystalChem) and a mouse adiponectin ELISA Kit (AssayPro). Liver enzymes were measured in plasma samples using an alanine aminotransferase (ALT) activity kit (DIO mice, Sigma-Aldrich) or ELISA Kit (NASH mice, Biomatik), a mouse aspartate aminotransferase (AST) ELISA Kit (Biomatik), a BCG (Bromocresol Green) albumin assay kit (Sigma-Aldrich), a glutamate dehydrogenase (GDH) activity assay kit (Sigma-Aldrich), and a bilirubin assay kit (Sigma-Aldrich).

Human primary hepatocytes were cultured in serum-free hepatocyte defined growth medium as described in section “Human primary hepatocyte culture”. Hepatocyte culture media was collected at indicated time points and stored at -20 °C before analysis. The concentration of urea was measured using urea assay kit and the concentration of albumin was assessed using BCG (Bromocresol Green) albumin assay kit (both Sigma-Aldrich) according to manufacturer’s instructions.