To remove excessive debris, the single cell suspension was passed through a 70-um filter into a 15 mL conical tube with 500 μl of high BSA buffer (ACSF with 1% FBS and 1% BSA) at the bottom to help cushion the cells during centrifugation at 100xg in a swinging bucket centrifuge for ...
After that time, the extract was centrifuged twice for 5 min in a 9.5 cm orbital radius centrifuge. The supernatant was transferred to a 25 mL volumetric flask, and made up to the mark with the same solvent. Finally, the extracts were stored at −20 ◦C prior to their analysis. ...