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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

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

Sigma catalog #

powder storage

concentration

amt for 25 mls

PMSF

P7626

room temp

100 mM

440 mg

compiled by Chad Rappleye

Aroian Lab Protocols