Protease Inhibitors
from Alberts' Lab Protocols (Kathy Miller/Chris Field)
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Protease inhibitors are rapidly inactivated in aqueous solutions and should be added to solutions immediately before use. Cocktail solutions are stored at -20 degrees and are good for approximately 6 months.
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I. Protease Inhibitor Cocktail
The Protease inhibitor cocktail is used as 100X for extract preps and as 1000X for column elution buffers**. Cocktail is made in 100% EtOH and contains:
inhbitor | ||||
Benzamidine HCL | ||||
Phenanthroline | ||||
Aprotinin* | ||||
Pepstatin A* | ||||
Leupeptin* |
* inhibitors purchased in one-time use amounts
** some components are not completely soluble in 100% EtOH. Vortex solution well before use to obtain a cloudy suspension and add to buffer - they become soluble in aqueous
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II. 100 mM PMSF
PMSF is very unstable in aqueous solutions. The 100 mM stock solution is made in isopropanol and stored in the dark at -20 degrees. Must be warmed (37 degrees) to get into solution. Use at 1 mM in solutions.
inhbitor | ||||
PMSF |