Thermo Scientific Halt Protease Inhibitor Cocktail, EDTA-free (100X) are ready-to-use concentrated stock solutions of protease inhibitors for addition to samples to prevent proteolytic degradation during cell lysis and protein extraction.
Features of Halt Protease Inhibitor Cocktail, EDTA-free (100X):
• Convenient—the refrigerator-stable, 100X liquid format is more effective and easier to use than individual inhibitors or other formats; just pipette exactly the amount you need • No proprietary ingredients—fully disclosed formulation contains optimized concentrations of six broad-spectrum protease inhibitors AEBSF, aprotinin, bestatin, E-64, leupeptin and pepstatin A stabilized in high-quality dimethylsulfoxide (DMSO) • Multiple package sizes—choose among several package sizes, including single-use microtubes that keep protease inhibitors fresh and reduce the risk of reagent contamination • EDTA-free formulations—ensures compatibility with isoelectric focusing • Lysis buffer compatible—use with Thermo Scientific Pierce Cell Lysis Buffers or nearly any other commercial or homemade detergent-based lysis reagent. Add 10 µL of concentrated cocktail per 1 mL of lysis buffer to ensure complete protection of the resulting protein extract.
Applications: • Protection of intact, active cellular proteins from degradation by endogenous proteases • Inhibition of protease activity from lysates produced by a variety of methods including Thermo Scientific Pierce Cell Lysis Reagents, other commercially formulated cell or tissue lysis products, sonication, French press, blender homogenization or repetitive freeze/thaw cycling • Screening of extracts for proteolytic activity • The study of proteolysis in the regulation of cellular processes
Proteases make up a large category of enzymes, including endopeptidases and exopeptidases, which catalyze the hydrolytic breakdown of proteins into peptides or amino acids in a wide variety of cell and tissue types. Proteases are regulated and compartmentalized in living cells, but cell lysis releases and mixes them with other extracted proteins. Their activity can thus lead to the loss of a large number of valuable proteins, adversely affecting downstream applications. For this reason, it is common practice to spike cell lysis buffers or cell extracts with a cocktails of compounds known to inactivate or inhibit proteases. Halt Protease Inhibitor Cocktails effectively inhibit serine-proteases, cysteine-proteases, aspartic acid-proteases and aminopeptidases that are typically present in cellular lysate samples. Regular and EDTA-free formulations provide for inhibition of metalloproteases and compatibility with 2D electrophoresis, respectively.