• »
• Lysis of Drosophila S2 Cells »
• Lysis of Drosophila S2 Cells in the Bullet Blender

Lysis of Drosophila S2 Cells in the Bullet Blender

Instrument: Bullet Blender or Bullet Blender Blue

Sample: Drosophila S2 Cells

Protocol:

1. Detach cells from culture dish or flask by your chosen method (trypsinization, scraping, spontaneous detachment, etc.)
2. Wash cells from dish with PBS into centrifuge tube
3. Centrifuge cell suspension to yield a cell pellet (1000x g for two minutes).
4. Pipette off the supernatant liquid.
5. Inspect the volume of the pellet. It should be 300μL or less in order to achieve efficient homogenization.
6. Add a volume of zirconium oxide beads (0.15mm) OR glass beads (0.1mm) to the tube equal to about ½ the volume of the pellet.
7. Add 2 volumes of buffer for every volume of cell pellet, minimum 25ml.
8. Close the microcentrifuge tubes.
9. Place tubes into the Bullet Blender®.
10. Set controls for SPEED 8 and TIME 2 minutes. Press Start.
11. After the run, remove tubes from the instrument and proceed with your downstream application.

Results:

Western blot with GAPDH stain from 72h culture of Drosophila S2 cells. Cells were homogenized per the above protocol, then the lysate was diluted 10x prior to loading in a 10% tris-glycine gel.

Data courtesy of Laura Stevens in the A. Page-McCaw lab at Rensselaer Polytechnic Institute