Silver: Lysate for Western
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- Spin down
- Wash with PBS
- Spin down enough cells to collect a 50-100 µL cell pellet in a FastPrep tube
-
PBSMT:
- 2.5 mM MgCl2
- 3 mM KCl
- 0.5% Triton X-100
- in PBS
-
Add protease inhibitors
-
PMSF: 1 mL per 100 mL buffer
- Use fresh PMSF: dissolve 0.035g per 1mL 100%EtOH
- PLAAC: 100 µL per 100 mL buffer
- 0.5 M Benzamidine: 260 µL per 100 mL buffer
-
PMSF: 1 mL per 100 mL buffer
- 6.5 m/s
- 40 second lysis
