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Protease Inhibitors

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  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.

 

compiled by Chad Rappleye

Aroian Lab Protocols

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