Protocol for Chloroform Methanol Precipitation

For Removal of salt and detergents

1. To sample of starting volume 100ul

2. Add 400ul methanol

3. Vortex well

4. Add 100ul chloroform

5. Vortex

6. Add 300ul H₂O

7. Vortex

8. Spin 1 minute @ 14,0000 g

9. Remove top aqueous layer (protein is between layers)

10. Add 400ul methanol

11. Vortex

12. Spin 2 minutes @ 14,000 g

13. Remove as much MeOH as possible without disturbing pellet

14. Speed-Vac to dryness

15. Bring up in 1X LSB

16. Boil 5 minutes

Reference:  Wessel, D. and Flugge, U. I. Anal. Biochem. (1984) 138, 141-143