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Protease and Phosphatase Inhibitors
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. Find the right protease, phosphatase, or combined liquid cocktails, tablets, and capsules for your experiments below.
Broad-spectrum protease inhibitor cocktails
| Liquid | Liquid, EDTA-free | Tablet | Tablet, EDTA-free | |
|---|---|---|---|---|
 |  |  |  | |
| Halt Protease Inhibitor Cocktail | Halt Protease Inhibitor Cocktail, EDTA-free | |||
| Contains EDTA | Yes, but in a separate vial | No | Yes | No |
| Contains DMSO | Yes | Yes | No | No |
| Requires reconstitution | No (100X concentrate) | No (100X concentrate) | Yes | Yes |
| Downstream compatibility | Not compatible with 2D or IMAC | All | Not compatible with 2D or IMAC | All |
| Included inhibitors | AEBSF, Aprotinin, Bestatin, E-64, Leupeptin, Pepstatin A, EDTA* | |||
| Cat. No. | 78430 (24 x 100 µL) 78429 (5 mL) 78438 (10 mL) 87786 (1 mL) | 78425 (24 x 100 µL) 78437 (5 mL) 78439 (10 mL) 87785 (1 mL) | A32963 (50 mL buffer per tablet) A32953 (Mini, 10 mL buffer per tablet) | A32965 (50 mL buffer per tablet) A32955 (Mini, 10 mL buffer per tablet) A37989 (XL, 500 mL buffer per capsule) |
*Only in EDTA formulations
Effective broad-spectrum protease inhibition
Figure 1. Pierce Protease Inhibitor tablets inhibits protease activity in different tissue lysates and cell lines. Protease activity of cell and tissue lysates (1 mg/mL) was determined in the presence (blue bars) and absence (black bars) of prepared Pierce Protease Inhibitor Tablet. The percentage of protease inhibition is indicated.
Strong inhibition of proteases
Click image to enlarge
Figure 2. 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 one hour at 37ºC and fluorescence was determined at the appropriate emissions. The percent protease inhibition is shown for each protease inhibitor formulation.
Broad-spectrum phosphatase inhibitor cocktails
| Liquid | Tablet | |
|---|---|---|
 | | |
| Halt Phosphatase Inhibitor Cocktail | Pierce Phosphatase Inhibitor Tablet | |
| Contains DMSO | Yes | No |
| Requires reconstitution | No (100X concentrate) | Yes |
| Downstream compatibility | Not compatible with 2D or IMAC | All |
| Included inhibitors | Sodium fluoride, sodium orthovanadate, sodium pyrophosphate, β-glycerophosphate | |
| Cat. No. | 78428 (24 x 100 uL) 78420 (1 mL) 78426 (5 x 1 mL) 78427 (10 mL) | A32957 (Mini, 10 mL buffer per tablet) |
Protecting protein phosphorylation status
Click image to enlarge
Figure 3. 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 mins 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 one hour 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 one hour at 37ºC and fluorescence was determined at the appropriate emissions. The percent phosphatase inhibition is shown for each phosphatase inhibitor formulation.
Inhibition of phosphatase activity to preserve phosphorylation state
Figure 4. Thermo Scientific Pierce Phosphatase Inhibitors preserve protein phosphorylation from phosphatases in cell and tissue extracts. Relative levels of total and phosphorylated protein were determined following protein extraction in the absence (–) or presence (+) of phosphatase inhibitors for (A) AKT and PDGFR in serum-starved, PDGF-stimulated (100 ng/mL) NIH 3T3 cell extracts and (B) ERK1/2 in liver and spleen tissue extracts. (C) The degree of inhibition for protein, acid, and alkaline phosphatase activity was determined in mouse brain extract following treatment with Pierce Phosphatase Inhibitor Tablets or another commercially available phosphatase inhibitor tablet.
Combined protease and phosphatase inhibitor cocktails
| Liquid | Liquid, EDTA-free | Tablet | Tablet, EDTA-free | |
|---|---|---|---|---|
| Halt Protease and Phosphatase Inhibitor Cocktail | Halt Protease and Phosphatase Inhibitor Cocktail, EDTA-free | Pierce Protease and Phosphatase Inhibitor Tablets | Pierce Protease and Phosphatase Inhibitor Tablets, EDTA-free | |
| Contains EDTA | Yes, but in a separate vial | No | Yes | No |
| Included inhibitors | Aprotinin, bestatin, E-64, leupeptin, sodium fluoride, sodium orthovanadate, sodium pyrophosphate, β-glycerophosphate, EDTA* | |||
| Contains DMSO | Yes | Yes | No | No |
| Requires reconstitution | No (100X concentrate) | No (100X concentrate) | Yes | Yes |
| Downstream compatibility | Not compatible with 2D or IMAC | All | Not compatible with 2D or IMAC | All |
| Cat. No. | 78442 (24 x 100 μL) 78440 (1 mL) 78444 (5 x 1 mL) 78446 (10 mL) | 78443 (24 x 100 μL) 78441 (1 mL) 78445 (5 x 1 mL) 78447 (10 mL) | A32959 (Mini, 10 mL buffer per tablet) | A32961 (Mini, 10 mL buffer per tablet) |
*Only in EDTA formulations
These combined protease and phosphatase inhibitor cocktails and tablets contain chemical compounds that target and inhibit serine, threonine, and tyrosine phosphatases, as well as serine, cysteine, and aspartic acid proteases, as well as aminopeptidases. Metalloproteases are inhibited by the addition of EDTA, which is available in a separate vial in the liquid format but included in the tablet format. These broad-spectrum inhibitor formulations prevent protein degradation and preserve phosphorylation simultaneously, helping provide complete protection in a single solution or tablet.
Figure 5. Effective inhibition of protease and phosphatase using the combination tablet. Using the tablet formulation that inhibits protease and phosphatase activity, the degree of enzyme inhibition was determined. A quenched-fluorescent substrate with pancreatic tissue extract was used and fluorescence measured after substrate cleavage. The degree of inhibition is indicated. (A) Protease assay reactions were incubated for two hours and 37°C. (B) Acid phosphatase assay reactions were incubated for one hour at 37°C.
Product Manuals
Protease Inhibitors
Phosphatase Inhibitors
Combined Protease and Phosphatase Inhibitors
Application notes
Broad-spectrum protease inhibitor cocktails
| Liquid | Liquid, EDTA-free | Tablet | Tablet, EDTA-free | |
|---|---|---|---|---|
| | | | |
| Halt Protease Inhibitor Cocktail | Halt Protease Inhibitor Cocktail, EDTA-free | |||
| Contains EDTA | Yes, but in a separate vial | No | Yes | No |
| Contains DMSO | Yes | Yes | No | No |
| Requires reconstitution | No (100X concentrate) | No (100X concentrate) | Yes | Yes |
| Downstream compatibility | Not compatible with 2D or IMAC | All | Not compatible with 2D or IMAC | All |
| Included inhibitors | AEBSF, Aprotinin, Bestatin, E-64, Leupeptin, Pepstatin A, EDTA* | |||
| Cat. No. | 78430 (24 x 100 µL) 78429 (5 mL) 78438 (10 mL) 87786 (1 mL) | 78425 (24 x 100 µL) 78437 (5 mL) 78439 (10 mL) 87785 (1 mL) | A32963 (50 mL buffer per tablet) A32953 (Mini, 10 mL buffer per tablet) | A32965 (50 mL buffer per tablet) A32955 (Mini, 10 mL buffer per tablet) A37989 (XL, 500 mL buffer per capsule) |
*Only in EDTA formulations
Effective broad-spectrum protease inhibition
Figure 1. Pierce Protease Inhibitor tablets inhibits protease activity in different tissue lysates and cell lines. Protease activity of cell and tissue lysates (1 mg/mL) was determined in the presence (blue bars) and absence (black bars) of prepared Pierce Protease Inhibitor Tablet. The percentage of protease inhibition is indicated.
Strong inhibition of proteases
Click image to enlarge
Figure 2. 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 one hour at 37ºC and fluorescence was determined at the appropriate emissions. The percent protease inhibition is shown for each protease inhibitor formulation.
Broad-spectrum phosphatase inhibitor cocktails
| Liquid | Tablet | |
|---|---|---|
| | |
| Halt Phosphatase Inhibitor Cocktail | Pierce Phosphatase Inhibitor Tablet | |
| Contains DMSO | Yes | No |
| Requires reconstitution | No (100X concentrate) | Yes |
| Downstream compatibility | Not compatible with 2D or IMAC | All |
| Included inhibitors | Sodium fluoride, sodium orthovanadate, sodium pyrophosphate, β-glycerophosphate | |
| Cat. No. | 78428 (24 x 100 uL) 78420 (1 mL) 78426 (5 x 1 mL) 78427 (10 mL) | A32957 (Mini, 10 mL buffer per tablet) |
Protecting protein phosphorylation status
Click image to enlarge
Figure 3. 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 mins 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 one hour 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 one hour at 37ºC and fluorescence was determined at the appropriate emissions. The percent phosphatase inhibition is shown for each phosphatase inhibitor formulation.
Inhibition of phosphatase activity to preserve phosphorylation state
Figure 4. Thermo Scientific Pierce Phosphatase Inhibitors preserve protein phosphorylation from phosphatases in cell and tissue extracts. Relative levels of total and phosphorylated protein were determined following protein extraction in the absence (–) or presence (+) of phosphatase inhibitors for (A) AKT and PDGFR in serum-starved, PDGF-stimulated (100 ng/mL) NIH 3T3 cell extracts and (B) ERK1/2 in liver and spleen tissue extracts. (C) The degree of inhibition for protein, acid, and alkaline phosphatase activity was determined in mouse brain extract following treatment with Pierce Phosphatase Inhibitor Tablets or another commercially available phosphatase inhibitor tablet.
Combined protease and phosphatase inhibitor cocktails
| Liquid | Liquid, EDTA-free | Tablet | Tablet, EDTA-free | |
|---|---|---|---|---|
| Halt Protease and Phosphatase Inhibitor Cocktail | Halt Protease and Phosphatase Inhibitor Cocktail, EDTA-free | Pierce Protease and Phosphatase Inhibitor Tablets | Pierce Protease and Phosphatase Inhibitor Tablets, EDTA-free | |
| Contains EDTA | Yes, but in a separate vial | No | Yes | No |
| Included inhibitors | Aprotinin, bestatin, E-64, leupeptin, sodium fluoride, sodium orthovanadate, sodium pyrophosphate, β-glycerophosphate, EDTA* | |||
| Contains DMSO | Yes | Yes | No | No |
| Requires reconstitution | No (100X concentrate) | No (100X concentrate) | Yes | Yes |
| Downstream compatibility | Not compatible with 2D or IMAC | All | Not compatible with 2D or IMAC | All |
| Cat. No. | 78442 (24 x 100 μL) 78440 (1 mL) 78444 (5 x 1 mL) 78446 (10 mL) | 78443 (24 x 100 μL) 78441 (1 mL) 78445 (5 x 1 mL) 78447 (10 mL) | A32959 (Mini, 10 mL buffer per tablet) | A32961 (Mini, 10 mL buffer per tablet) |
*Only in EDTA formulations
These combined protease and phosphatase inhibitor cocktails and tablets contain chemical compounds that target and inhibit serine, threonine, and tyrosine phosphatases, as well as serine, cysteine, and aspartic acid proteases, as well as aminopeptidases. Metalloproteases are inhibited by the addition of EDTA, which is available in a separate vial in the liquid format but included in the tablet format. These broad-spectrum inhibitor formulations prevent protein degradation and preserve phosphorylation simultaneously, helping provide complete protection in a single solution or tablet.
Figure 5. Effective inhibition of protease and phosphatase using the combination tablet. Using the tablet formulation that inhibits protease and phosphatase activity, the degree of enzyme inhibition was determined. A quenched-fluorescent substrate with pancreatic tissue extract was used and fluorescence measured after substrate cleavage. The degree of inhibition is indicated. (A) Protease assay reactions were incubated for two hours and 37°C. (B) Acid phosphatase assay reactions were incubated for one hour at 37°C.
Product Manuals
Protease Inhibitors
Phosphatase Inhibitors
Combined Protease and Phosphatase Inhibitors
Application notes
Related products and downstream applications
For Research Use Only. Not for use in diagnostic procedures.