Animal cell protein extraction for gel-free protein expression study
1. Add 5 ml of PBS buffer to the cell and briefly vortex
2. Centrifuge the suspension at 1,000 rpm for 5 min at room temperature
3. Remove the supernatant with a pipette
4. Add 5 ml of PBS buffer the cell and briefly vortex
5. Centrifuge the suspension at 1,000 rpm for 5 min at room temperature
6. Remove the supernatant with a pipette
7. Add 5 ml of PBS buffer the cell and briefly vortex
8. Centrifuge the suspension at 1,000 rpm for 5 min at room temperature
9. Remove the supernatant with a pipette
10. Add 0.1% RapiGest (Waters, Catalog # 186001861) (100µl per 1,000,000 cells) to the cells. Incubate 30 min at 22 ºC. (If you have Bead-beater, you can use Bead- Beater in this step).
11. Centrifuge the suspension at 14,000 x g for 10 min at room temperature
12. Collect and save the supernatant
13. Total protein test
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