1.1.2 Mammalian Suspension Cells

Operational Steps for Protein Extraction from Mammalian Suspension Cells

1.Collect Cells
Transfer the culture medium in the culture flask to a 15 ml centrifuge tube, and centrifuge at 1000 rpm for 4 minutes to collect the cell pellet.
2.Rinse
Add pre-cooled PBS to transfer the cell pellet to a 1.5 ml EP tube, centrifuge and rinse with pre-cooled PBS for 3 times, and finally aspirate the PBS.
3.Lysis
Add 500 μl of cell lysis buffer (note: protease and phosphatase inhibitors must be added to the lysis buffer in advance), and lyse on ice for 10–20 minutes.
4.Centrifuge to Collect Supernatant
Centrifuge at 12,000 rpm at 4°C for 5 minutes. Collect the supernatant, aliquot it, and store at -20°C or -80°C for later use.