Protease Inhibitor Cocktail solution (mammalian cell and tissue extract use) (EDTA free)

Overview

This protease inhibitor cocktail contains a mixture of protease inhibitors with broad specificity for various proteases. It is optimized for use in mammalian cell and tissue extracts. Protease inhibitor cocktails protect proteins from degradation by endogenous proteases released during protein extraction and purification. This cocktail is EDTA free.

Components and action

The following components are included in this cocktail which target the major classes of proteases including serine, cysteine and acid proteases and also aminopeptidases.

• AEBSF hydrochloride serine protease inhibitor (100mM)
• Aprotinin (bovine recombinant, Nicotiana sp.,): serine protease inhibitor (80µM)
• Bestatin: aminopeptidase B and leucine aminopeptidase inhibitor (5mM)
• E-64: cysteine protease inhibitor (1.5mM)
• Leupeptin hemisulfate: serine/cysteine protease inhibitor (2mM)
• Pepstatin A: aspartic protease inhibitor (1mM)

Formulation & usage recommendation

The cocktail is supplied as a 1ml solution of 100X concentrate in DMSO. It is generally effective at a 1X final concentration but may require optimization if a sample contains particularly high levels of proteases.

1mL of this cocktail solution is recommended for inhibition of endogenous enzymes found in 100 mL of lysate from 20g of bovine liver or 10 mL of lysate from CHO cells. Please note that not all lysates contain the same levels of endogenous enzymes and you may need to adjust the volume of cocktail required accordingly.

1. Equilibrate the bottle to room temperature.
2. Vortex the bottle to ensure a homogeneous suspension.
3. Immediately before use, add 10µL of the Protease Inhibitor Cocktail per milliliter of sample directly to the lysis buffer or extract to produce a 1X final concentration.