5.3. Measurement of alkaline phosphatase activity

ALP activity was measured using a commercially obtained assay kit (Abcam, Cambridge, UK) following the manufacturer's instructions. Briefly, jejunum explant lysates and hepatocyte lysates were incubated with para‐nitrophenyl phosphate (pNPP) substrate for 1 hr at 25°C, following which the absorbance was measured at 405 nm to quantify the para‐nitrophenol (pNP) produced through ALP‐mediated dephosphorylation. A standard curve was generated using the pNPP substrate and ALP enzyme provided by the manufacturer.

where A = concentration of pNP generated in samples (μmol), V = volume of sample (ml), T = reaction time (min). ALP activity values were normalized to the protein content of each sample.