4.16. Alkaline Phosphatase (ALP) Activity Assay

To measure the impact of fucoidan in MSC mono-cultures on ALP activity, the supernatant was collected 7 days after treating with 1, 10, and 100 µg/mL fucoidan, and an alkaline phosphatase assessment was performed according to the manufacturer’s protocol from the kit (Abcam, Germany). In brief, samples and standards were prepared in triplicates in 96-well plates for ALP reaction with pNPP Solution. Stop solution was added to the control wells before starting the ALP reaction to determine the background values. After 60 min incubation, the reaction in samples was stopped by stop solution, and the OD value was measured at 405 nm with a microplate reader. The values were calculated after background subtraction and are presented relative to the control.