Protease Inhibitor Cocktail (EDTA-Free, 100X in DMSO)

Protease inhibitor cocktails (similar to Sigma P8849) are commonly used in protein extraction to prevent protein breakdown by natural cellular proteases. These enzymes (including cysteine, serine, acid proteases, and aminopeptidases) are activated when cells or tissues are disrupted and can damage target proteins. This ready-to-use 100X DMSO solution uses a mix of inhibitors (AEBSF, Bestatin, E-64, Leupeptin, Pepstatin A) to block such proteases, preserving proteins’ natural structure and properties. Being EDTA-free, it works well for downstream steps sensitive to divalent cations, like phosphorylation analysis or enzyme assays.

Cellular crude extracts contain numerous endogenous enzymes, such as phosphatases and proteases, capable of degrading proteins in the extracts. The most effective strategy to enhance the yield of intact proteins is through the addition of inhibitors targeting these known endogenous enzymes.The Protease Inhibitor Cocktail is intended for the purification of His-tagged proteins to improve protein stability. This cocktail functions by inhibiting proteases that degrade either phosphorylated or non-phosphorylated protein substrates.

This Protease Inhibitor Cocktail contains individual components including AEBSF, Bestatin, E-64, Phosphoramidon, and Pepstatin A, offering broad-spectrum specific inhibition against cysteine, serine, aspartic, thermolysin-like proteases, and aminopeptidases. It has been optimized and validated specifically for use with His-tagged proteins. The cocktail is dissolved in DMSO and supplied as a ready-to-use solution.