Protease Inhibitor Cocktail (100X)

Protease Inhibitor Cocktail (ab271306) protects protein extracts from aminopeptidases, metalloproteases, and serine, cysteine, and aspartic acid proteases. It is supplied as a 100X stock solution of protease inhibitors dissolved in DMSO and a 100X stock solution of EDTA dissolved in water, pH 8.0. ab271306 is a general-purpose reagent suitable for protease inhibition during protein extraction from mammalian tissues and cells, yeast and fungal cells, and bacteria.

ab271306 contains the chelating agent EDTA for improved protection against metalloproteases. It should therefore not be added to samples intended for subsequent analysis by techniques that are incompatible with EDTA, such as isoelectric focusing and metal affinity column separations. For these applications, use Protease Inhibitor Cocktail EDTA Free (Protease Inhibitor Cocktail EDTA Free (100X) ab270055).

This product is manufactured by Expedeon, an Abcam company. It was previously called Proteoloc™ Protease Inhibitor Cocktail. Expedeon product code 42506 is the same as the 3 mL size of this product, 42507 is the same as the 30 mL size, and 44204 is the same as the 10 mL size.

Composition:

AEBSF: Irreversible serine protease inhibitor. Inhibits chymotrypsin, kallikrein, plasmin, thrombin, and trypsin.

Aprotinin: Reversible serine protease inhibitor. Inhibits chymotrypsin, kallikrein, plasmin, trypsinogen, and urokinase.

Bestatin: Metalloprotease inhibitor selective for aminopeptidases. Inhibits activity of leucine aminopeptidase, aminopeptidase B, and triamino peptidase.

EDTA: Inhibits metalloproteases by chelating divalent metal ions (provided in a separate vial).

E-64: Irreversible inhibitor of cystine proteases. Inhibits calpain, papain, and cathespin B.

Leupeptin: Reversible inhibitor of serine and cystine proteases. Inhibits calpain, papain, trypsin, and cathespin B.

Pepstatin A: Reversible aspartic acid protease inhibitor. Inhibits pepsin, rennin, cathespin D, chymosin, and protease B.

Recommended procedure:

Allow product to thaw at room temperature. Agitate or vortex container to ensure uniform solution. ab271306 is provided as a 100X stock solution of protease inhibitors and a 100X stock solution of EDTA. Dilute product into desired lysis buffer to obtain a 1X concentration (5 mM). For example, dilute 100 μL protease inhibitors and 100 μL EDTA into 10 mL Protein Extraction Kit (Protein Extraction Kit ab270054). Proceed in accordance with extraction kit or lysis buffer protocol.