蛋白酶抑制剂资料 Protease Inhibitors
Protease Inhibitors
from Alberts' Lab Protocols (Kathy Miller/Chris Field)
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.
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 | Sigma catalog # | powder storage | concentration in cocktail | amt for 25 mls cocktail |
Benzamidine HCL | B6506 | 4 deg C | 1.6 mg/ml | 40 mg |
Phenanthroline | P1294 | room temp | 1 mg/ml | 25 mg |
Aprotinin* | A1153 | 4 deg C | 1 mg/ml | 25 mg |
Pepstatin A* | P5318 | 4 deg C | 1 mg/ml | 25 mg |
Leupeptin* | L2884 | -20 deg C | 1 mg/ml | 25 mg |
* 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
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.
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