Harvesting of Cells For Protein Extraction in Cos 7 Cells

1. Decant Medium
2. Add 10-20ml of 1x PBS Swirl.Decant. Repeat.
3. Add 10ml of 1x PBS. Remove cell layer by scraping with rubber policeman.
4. Collect the PBS/cells in 15ml centrifuge tube. Spin 5 min at 4C (1000pm).
5. ACarefully decant solution, leaving about 100ul of PBS
6. Transfer cells to microcentrifuge tube by resuspending cells in remaining PBS.
7. Spin as above.
8. Freeze @ -80C or continue...
9. Resuspend in 150ul PBS and 1.5ul PI1 (Protease Inhibitor Mix)
10. Sonicate
11. Spin top speed, 3 min and collect supernatant.
12. Freeze @ -80C.
    note: may also add PMSF, MC (Phos.Inhib), Beta-glycerophospate.