Protease and Phosphatase Inhibition

Protease and phosphatase inhibitor cocktails, tablets and capsules are ideal for the protection of proteins during extraction or lysate preparation from primary cells, cultured mammalian cells, animal tissues, plant tissues, yeast or bacterial cells. Individual protease inhibitors are also available separately and in multiple sizes. Formulations with or without EDTA are available for divalent cation-sensitive assays. Learn more about our protease, phosphatase or combined liquid cocktails, tablets and capsules.

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  • Effective—outperforms leading competitor formulations across targeted protease and phosphatase families
  • Multiple package sizes—liquid cocktails, tablets and capsules are available in multiple sizes and formats to accommodate different volume and pricing needs
  • Improved formulation—tablets and capsule contents dissolve quickly into a clear solution, and are fully compatible with all Pierce protein assays.
  • Convenient—refrigerator-stable formulations are easier to use than individual inhibitors
  • Complete protection—all-in-one formulations contain both protease and phosphatase inhibitors (combined cocktail only)
  • Compatible—does not interfere with protein assays or other downstream applications

Choose a broad spectrum cocktail or individual inhibitor

Inhibitor component Target (mechanism) Protease liquid cocktails and tablets Protease capsules Phosphatase liquid cocktails and tablets Combined protease and phosphatase liquid cocktails and tablets
AEBSF∙HCL Serine protease (irreversible)

   
Aprotinin Serine protease (irreversible)

   

Bestatin Aminopeptidase (reversible)

 

E-64 Cysteine (irreversible)

 

Leupeptin Serine and cysteine protease (reversible)

   

Pepstatin Aspartic acid proteases (reversible)

   
PMSF Serine proteases (irreversible)        
EDTA†* Metalloproteases

   

Sodium fluoride* Serine-threonine and acidic phosphatases    

Sodium orthovanadate* Tyrosine and alkaline phosphatases    

Β-glycero-phosphate* Serine-threonine phosphatases    

Sodium pyrophosphate* Serine-threonine phosphatases    

†Not in EDTA-free formulations
*Not available individually


Compare protease inhibitor cocktails and tablets

Performance comparison between three commercially available protease inhibitor tablets

Figure 1. Performance comparison between three commercially available protease inhibitor tablets. Pancreatic extract (100 μL; 0.5 μg/μL) was incubated with quenched, fluorescent protease-cleavable substrates for trypsin, cysteine, and metalloprotease and cathepsins in the presence of the reformulated Thermo Scientific Pierce Protease Inhibitor Mini Tablets, Roche™ cOmplete™ Protease Inhibitor Tablets, and Sigma-Aldrich™ SIGMAFAST™ Protease Inhibitor Cocktail Tablets with and without EDTA. Reactions were incubated for 1 hr at 37ºC and fluorescence was determined at the appropriate emissions. The percent protease inhibition is shown for each protease inhibitor formulation.

 Protein phosphorylation is preserved in cell extracts

Figure 2. Protein phosphorylation is preserved in cell extracts. Relative levels of total and phosphorylated protein from extracts prepared in the absence or presence of phosphatase inhibitors were determined by western blot analysis. (A) HCT116 cells were serum-starved and treated with either IGF for 15 min or left as control cells. Cell lysates were prepared in IP Lysis Buffer and the reformulated Thermo Scientific protease and phosphatase combined inhibitors (EDTA-free). 500 μg of lysate was incubated with 5 μg of phospho-AKT antibody overnight at 4ºC. The complex was then incubated with Thermo Scientific Pierce Protein A/G Magnetic beads for 1 hr at RT. Beads were washed and low-pH elution was performed. (B) The degree of inhibition for protein, alkaline, and acid phosphatase activity was determined in kidney extract (25 μL; 0.5 μg/μL) by incubating extracts with a fluorogenic substrate (MFP or FDP) that measures phosphatase activity upon dephosphorylation in the presence of Pierce Phosphatase Inhibitor Mini Tablets, Roche™ PhosStop™ Phosphatase Inhibitor Tablets, and Sigma-Aldrich™ Phosphatase Inhibitor Cocktail 2 and 3 liquid formulations. Reactions were incubated for 1 hour at 37ºC and fluorescence was determined at the appropriate emissions. The percent phosphatase inhibition is shown for each phosphatase inhibitor formulation.