Microbial cell protein extraction for gel electrophoresis
1. Add 1 ml of PBS buffer to the cell and briefly vortex
2. Centrifuge the suspension at 8,000 rpm for 4 min at room temperature
3. Remove the supernatant with a pipette
4. Add 1 ml of PBS buffer the cell and briefly vortex
5. Centrifuge the suspension at 8,000 rpm for 4 min at room temperature
6. Remove the supernatant with a pipette
7. Add 1 ml of PBS buffer the cell and briefly vortex
8. Centrifuge the suspension at 8,000 rpm for 4 min at room temperature
9. Remove the supernatant with a pipette
10. Add Lysis buffer (Tris 30 mM, Urea 8 M, CHAPS 4%) with inhibitor (~100µl per 100,000,000 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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