Thermo Scientific Pierce Protease Inhibitor Mini Tablets, EDTA-free, contain broad-spectrum protease inhibitors that are highly effective at preventing proteolytic degradation during cell lysis and protein extraction experiments. Pierce Protease Inhibitor Mini Tablets protect proteins from proteolytic degradation through inhibition of the serine proteases, cysteine proteases, aspartic acid proteases, and aminopeptidases that are typically present in cell lysate samples. The tablets contain AEBSF, aprotinin, bestatin, E-64, leupeptin, and pepstatin A. Stable for storage at 4°C for up to 12 months, each mini tablet is sufficient for 10 mL of solution, and is suitable for reagent preparation prior to protein extraction from tissue and cultured cells. Proteases make up a large category of enzymes, including endopeptidases and exopeptidases, which catalyze the hydrolytic breakdown of proteins into peptides or amino acids. Proteases reside in cellular compartments, but upon cell lysis the proteases are released, resulting in the degradation of valuable proteins. For this reason, it is common practice to add multiple compounds known to inactivate or inhibit proteases to cell lysis buffers or cell extracts. To use Pierce Protease Inhibitor Mini Tablets, simply dissolve 1 tablet in 10 mL of buffer or lysate. The protease inhibitor formulation is compatible with most detergent-based cell lysis reagents, including Pierce Cell Lysis solutions. Pierce Protease Inhibitor Mini Tablets do not contain phosphatase inhibitors. If necessary, phosphatase inhibitors, such as Pierce Phosphatase Inhibitor Mini Tablets, can be added along with the protease inhibitor tablets. |
图1 Comparison of commercially-available protease inhibitor tablets Pancreatic extract (100 µL; 0.5 µg/µL) was incubated with fluorescent protease-cleavable substrates for trypsin, cysteine, and metalloprotease/cathepsins in the presence of commercially-available protease inhibitors with and without EDTA. Reactions were incubated for 1 hour at 37ºC and fluorescence was measured. The percent protease inhibition is shown for each protease inhibitor formulation. |