Protease Inhibitor Cocktail

Molecular Biology / MB-Protease Inhibitors

-20°C

Supplied as lyophilized powder containing 2mM AEBSF, 1mM EDTA, 130uM Bestatin, 14uM E-64, 1uM Leupeptin, 0.3uM Aprotinin when reconstituted in 100ml of buffer.

Applications:
Used generally to inhibit proteases (serine, cysteine, aspartic and metalloproteases).

One bottle is sufficient to inhibit proteases present in 20g of cellular extract. Note that some organisms contain higher levels of proteases and may require a higher concentration of protease cocktail to inhibit the intrinsic proteolytic activity.

Activity:
One ml of the cocktail will inhibit proteolytic activity equivalent to 1mg of USP pancreatin.

Inhibitor Targets:
AEBSF: Serine proteases, such as trypsin and chymotrypsin
EDTA Metalloproteases
Bestatin: Aminopeptidases, such as leucine aminopeptidase and alanyl aminopeptidase
E-64: Cysteine and thiol proteases, such as calpain, papain, cathepsin B and cathespin L
Leupeptin: Serine and thiol proteases, such as calpain, trypsin, papain and cathepsin B Aprotinin: Serine proteases such as trypsin, chymotrypsin, plasmin, trypsinogen, urokinase and kallikrein. Inhibits leukocyte elastase but not pancreatic elastase

Reconstitution:
Dissolve powder in 10ml of RIPA lysis buffer (10X) and transfer to a larger container to prepare 100ml final volume. The powder may be prepared and stored as a more concentrated stock for further dilution.

Storage and Stability:
Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute with sterile buffer. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 1 week at 4ºC.