Protease Inhibitor Cocktail (EDTA-Free, 100X in H2O)

Endogenous proteins are produced and degraded in equilibrium and therefore their cellular level is stable under stable environmental conditions. Crude cell extracts contain many endogenous enzymes capable of degrading proteins in the extract, such as phosphatases and proteases. When proteins are extracted from cells and tissues in vitro, protein production is significantly inhibited, and degradation increases. The best way to increase intact protein production is to add inhibitors of those enzymes that are known to be present. The protease inhibitor cocktail is used in cell lysates or tissue extracts to improve protein stability. 
The product is a ready-to-use water-soluble mixed concentrate containing EDTA, which has broad specificity and can be used as a spectral protease inhibitor mixture. The protease inhibitor cocktail contains several important components, including AEBSF, Aprotinin, Bestatin hydrochloride, E-64, and Leupeptin, which are broadly specific for serine, cysteine, acid proteases, and aminopeptidases. The product can be used for Western Blot, Co-IP, pull-down, IF, IHC, Kinase assay and other experiments.
It is important to note that EDTA inhibits enzyme activity by chelating divalent cations necessary for metalloproteinase activity, so EDTA may affect the activity of other proteins. Pre-experiments may be required in specific experiments to determine whether EDTA has adverse effects. If the protein of interest is to be purified using immobilized metal chelation affinity chromatography (IMAC) or 2D gel electrophoresis analysis is required, EDTA must be removed by extensive dialysis or desalting prior to purification.
This product is supplied as a ready-to-use concentrated solution (100×, in H₂O) with a size of 1 mL. For additional metalloprotease inhibitor components such as EDTA, Cat. No. K4003 can be used alternatively.