Application

Protease Inhibitor Cocktail has been used-as a buffer component during sonication of GFP (green fluorescent protein)-huntingtin-transfected HEK 293 cells in GST (glutathione S-transferase) pull down assayas a component of lysis bufferas a component of radioimmunoprecipitation assay buffer (RIPA)

Biochem/physiol Actions

This mixture contains individual components, including AEBSF at 104 mM, Aprotinin at 80 µM, Bestatin at 4 mM, E-64 at 1.4 mM, Leupeptin at 2 mM and Pepstatin A at 1.5 mM. Each component has specific inhibitory properties. AEBSF and Aprotinin act to inhibit serine proteases, including trypsin, chymotrypsin, and plasmin amongst others. Bestatin inhibits aminpeptidases. E-64 acts against cystein proteases. Leupeptin acts against both serine and cystein proteases. Pepstatin A inhibits acid proteases.

Caution

This cocktail should be stored at -20 °C.

Components

AEBSFAprotininBestatinE-64LeupeptinPepstatin A

Packaging

1, 5 mL in serum bottle

Preparation Note

One mL of this cocktail solution is recommended for the inhibition of endogenous enzymes found in 100 mL of lysate from 20 g of bovine liver or 10 mL of lysate from CHO cells. This product is supplied as a solution in DMSO.

Specificity

Inhibits serine, cysteine, and acid proteases, and aminopeptidases.