Search Thermo Fisher Scientific
Protease and Phosphatase Inhibitors |
Protease and phosphatase inhibitors are essential components in biochemical research and protein studies as they safeguard the structural integrity of proteins during the extraction and analysis processes. Protease inhibitors act by blocking protease activity, thus helping ensure proteins remain in their native form throughout various experimental procedures. Similarly, phosphatase inhibitors prevent the dephosphorylation of proteins, crucial for maintaining their phosphorylation status. This is particularly vital for studying signaling pathways, protein-protein interactions, and other cellular processes where phosphorylation plays a critical role.
During protein extraction, it is important to inhibit the activity of these enzymes to prevent degradation and alteration of protein samples. By preserving the native state and functionality of proteins, researchers can obtain accurate, reliable, and reproducible results from their experimental studies.
Thermo Fisher Scientific offers a variety of protease inhibitors, phosphatase inhibitors, and combined inhibitor cocktails that effectively inhibit against a broad spectrum of proteases and phosphatases. These are available in different formats—cocktails, tablets, and capsules—and with or without EDTA. Choose from our comprehensive range of protease inhibitors and/or phosphatase inhibitors based on the specific needs of your experiment.
Learn more: Protease and phosphatase inhibitors
On this page:
Related content
Choose your optimal protease and phosphatase inhibitor
Protease inhibitors prevent proteolytic degradation during cell lysis and protein extraction. These inhibitors contain cysteine protease inhibitors (E-64), serine protease inhibitors (AEBSF, Aprotinin, Leupeptin), and metalloprotease inhibitors (Bestatin, Pepstatin A, EDTA*)
 |  |  |  | ||||||
| Halt Protease Inhibitor Cocktail | Halt Protease Inhibitor Cocktail, EDTA-free | Pierce Protease Inhibitor tablet | Pierce Protease Inhibitor mini tablet | Pierce Protease Inhibitor tablets, EDTA-free | Pierce Protease Inhibitor mini tablets, EDTA-free | Pierce Protease Inhibitor XL Capsules, EDTA-free | |||
| Format | Liquid | Liquid, EDTA-free | Tablet | Tablet, 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 | |||||
| Available sizes | 24 x 100 µL 1 mL 5 mL 10 mL | 24 x 100 µL 1 mL 5 mL 10 mL | 50 mL buffer per tablet | Mini, 10 mL buffer per tablet | 50 mL buffer per tablet | Mini, 10 mL buffer per tablet | XL, 500 mL buffer per capsule | ||
| Documents | User Guide: Halt Protease Inhibitor Cocktail | Product Sheet: Pierce Protease Inhibitor Tablets | Product Sheet: Pierce Protease Inhibitor XL Capsules, EDTA-Free | ||||||
*only in EDTA formulations
Broad-spectrum protease inhibitors
Protease inhibitors are available as a Halt Protease Inhibitor Cocktail and Pierce Protease Inhibitor tablet formats. Both formats inhibit serine-proteases, cysteine-proteases, aspartic acid-proteases, and aminopeptidases (Table 1). Metalloproteases are inhibited by the addition of EDTA, which is available in a separate vial in the cocktail format but included in the tablet format. Protease inhibitors can be added to cell lysis buffers or cell extracts to prevent protein degradation.
Pierce Protease Inhibitor tablets have an inhibitory effect on a broad range of proteases in various tissues and cell types (Figure 1). Protease inhibitor tablets are easy-to-use, simply dissolve 1 tablet in 50 mL of buffer or lysate or 1 mini tablet in 10 mL of buffer or lysate. For Protease Inhibitor XL capsules, dissolve 1 capsule in 500 mL of buffer.
Halt Protease Inhibitor Cocktails are compatible with nearly any commercially available cell lysis buffer or homemade lysis reagent. Simply add 100 µL of the concentrated inhibitor to 1 mL of lysis buffer. Protease inhibitor cocktails have been shown to have significant protease inhibition compared to commercially available tablet products (Figure 2).
EDTA-free protease inhibitors are also available, as EDTA can affect some protein activity and interfere with immobilized metal-chelate affinity chromatography (IMAC) and 2D gel electrophoresis.
Table 1. Formulation and concentration of the Thermo Scientific Halt Protease Inhibitor Cocktail.
Click image to enlarge
Figure 1. Pierce Protease Inhibitor tablets inhibit 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.
Click image to enlarge
Figure 2. Performance comparison of commercial protease inhibitor cocktails and tablets. Using a validated protease assay and 1.0mg/mL of rat pancreas extract, the Thermo Scientific Halt Protease Inhibitor Single-Use Cocktails were tested against other commercially available tablet-format protease inhibitor cocktails under the same conditions. A 1X final concentration of each inhibitor was added. The single-use formulation resulted in ≥97% inhibition compared to ≥59% inhibition for the tablet.
Phosphatase inhibitors prevent the dephosphorylation of proteins, maintaining their phosphorylation status. These inhibitors contain sodium fluoride, sodium orthovanadate, sodium pyrophosphate, and β-glycerophosphate.
 | | |
| Halt Phosphatase Inhibitor Cocktail | Pierce Phosphatase Inhibitor Tablet | |
| Format | Liquid | Tablet |
| Contains DMSO | Yes | No |
| Requires reconstitution | No (100X concentrate) | Yes |
| Downstream compatibility | Not compatible with 2D or IMAC | All |
| Available sizes | 24 x 100 uL 5 x 1 mL 1 mL 10 mL | Mini, 10 mL buffer per tablet |
| Documents | User Guide: Halt Phosphatase Inhibitor Cocktail | Product Sheet: Pierce Phosphatase Inhibitor Mini Tablets |
Broad-spectrum phosphatase inhibitors
Phosphatase inhibitors are available as a Halt Phosphatase Inhibitor Cocktail and Pierce Phosphatase Inhibitor Mini tablet formats. Both formats contain sodium fluoride, sodium orthovanadate, sodium pyrophosphate and beta-glycerophosphate inhibitors. Phosphatase inhibitors are compatible with cell lysis reagents.
The Halt Phosphatase Inhibitor Cocktail preserves the phosphorylation state of proteins by inhibiting serine/threonine phosphatases and tyrosine phosphatases during cell lysis or tissue protein extraction (Figure 3). It should be noted that the Halt phosphatase inhibitor cocktail interferes with 2D gel electrophoresis, and some immobilized metal chelate affinity chromatography methods but is compatible with the Pierce Phosphoprotein Enrichment Kit. Before performing incompatible procedures, either dialyze or desalt the sample to remove inhibitors from extracts.
The Pierce Phosphatase Inhibitor Mini tablets protect phosphorylation from serine/threonine phosphatases. The Pierce Phosphatase Inhibitor Mini tablet is suitable for use with any cell or tissue extract (Figure 4). To use, simply dissolve 1 phosphatase inhibitor mini tablet in 10 mL of solution.
Click image to enlarge
Figure 3. Preserved phosphorylation states of three proteins in cell lysate. HeLa cells were lysed in the absence (-) and presence (+) of a 1X concentration of the Thermo Scientific Halt Phosphatase Inhibitor Cocktail. Cell lysates were analyzed by Western blot for total and phosphorylated protein as indicated. Panel A: MEK and phosphorylated MEK (PMEK). Panel B: MAP Kinase and PMAP Kinase. Panel C: STAT3 and PSTAT3. The proteins are phosphorylated on serine, threonine/tyrosine and tyrosine, respectively.
Click image to enlarge
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 wothout inhibitor.
Protease and phosphatase cocktails preserve the native state of proteins and their post-translational modifications, helping ensure samples remain stable. These inhibitors contain cysteine protease inhibitors (E-64), serine protease inhibitors (Aprotinin, Leupeptin), and metalloprotease inhibitors (Bestatin, EDTA*) along with phosphatase inhibitors (sodium fluoride, sodium orthovanadate, sodium pyrophosphate, β-glycerophosphate).
 |  |  | ||
| 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 | |
| Format | Liquid | Liquid, EDTA-free | Tablet | Tablet, 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 |
| Available sizes | 24 x 100 uL 5 x 1 mL 1 mL 10 mL | 24 x 100 uL 5 x 1 mL 1 mL 10 mL | Mini, 10 mL buffer per tablet | Mini, 10 mL buffer per tablet |
| Documents | Product Sheet: Halt Protease and Phosphatase Inhibitor Cocktail | Product Sheet: Pierce Protease and Phosphatase Inhibitor Mini Tablets | ||
*only in EDTA formulations
Combined protease and phosphatase inhibitors
Combined Protease and Phosphatase Inhibitors are available in cocktail and tablet formats. These broad-spectrum inhibitors protect against protein degradation while preserving protein phosphorylation simultaneously. They contain chemical compounds that inhibit serine-, cysteine-, and aspartic acid-proteases, and serine/threonine and tyrosine phosphatases, as well as aminopeptidases (Table 2). Metalloproteases are inhibited by the addition of EDTA, which is available in a separate vial in the cocktail format but included in the tablet format (Figure 5).
Table 2. Protease and phosphatase inhibitors included in the cocktail formulation and their targeted enzyme class.
Click image to enlarge
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.
Protease inhibitors prevent proteolytic degradation during cell lysis and protein extraction. These inhibitors contain cysteine protease inhibitors (E-64), serine protease inhibitors (AEBSF, Aprotinin, Leupeptin), and metalloprotease inhibitors (Bestatin, Pepstatin A, EDTA*)
| | | | ||||||
| Halt Protease Inhibitor Cocktail | Halt Protease Inhibitor Cocktail, EDTA-free | Pierce Protease Inhibitor tablet | Pierce Protease Inhibitor mini tablet | Pierce Protease Inhibitor tablets, EDTA-free | Pierce Protease Inhibitor mini tablets, EDTA-free | Pierce Protease Inhibitor XL Capsules, EDTA-free | |||
| Format | Liquid | Liquid, EDTA-free | Tablet | Tablet, 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 | |||||
| Available sizes | 24 x 100 µL 1 mL 5 mL 10 mL | 24 x 100 µL 1 mL 5 mL 10 mL | 50 mL buffer per tablet | Mini, 10 mL buffer per tablet | 50 mL buffer per tablet | Mini, 10 mL buffer per tablet | XL, 500 mL buffer per capsule | ||
| Documents | User Guide: Halt Protease Inhibitor Cocktail | Product Sheet: Pierce Protease Inhibitor Tablets | Product Sheet: Pierce Protease Inhibitor XL Capsules, EDTA-Free | ||||||
*only in EDTA formulations
Broad-spectrum protease inhibitors
Protease inhibitors are available as a Halt Protease Inhibitor Cocktail and Pierce Protease Inhibitor tablet formats. Both formats inhibit serine-proteases, cysteine-proteases, aspartic acid-proteases, and aminopeptidases (Table 1). Metalloproteases are inhibited by the addition of EDTA, which is available in a separate vial in the cocktail format but included in the tablet format. Protease inhibitors can be added to cell lysis buffers or cell extracts to prevent protein degradation.
Pierce Protease Inhibitor tablets have an inhibitory effect on a broad range of proteases in various tissues and cell types (Figure 1). Protease inhibitor tablets are easy-to-use, simply dissolve 1 tablet in 50 mL of buffer or lysate or 1 mini tablet in 10 mL of buffer or lysate. For Protease Inhibitor XL capsules, dissolve 1 capsule in 500 mL of buffer.
Halt Protease Inhibitor Cocktails are compatible with nearly any commercially available cell lysis buffer or homemade lysis reagent. Simply add 100 µL of the concentrated inhibitor to 1 mL of lysis buffer. Protease inhibitor cocktails have been shown to have significant protease inhibition compared to commercially available tablet products (Figure 2).
EDTA-free protease inhibitors are also available, as EDTA can affect some protein activity and interfere with immobilized metal-chelate affinity chromatography (IMAC) and 2D gel electrophoresis.
Table 1. Formulation and concentration of the Thermo Scientific Halt Protease Inhibitor Cocktail.
Click image to enlarge
Figure 1. Pierce Protease Inhibitor tablets inhibit 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.
Click image to enlarge
Figure 2. Performance comparison of commercial protease inhibitor cocktails and tablets. Using a validated protease assay and 1.0mg/mL of rat pancreas extract, the Thermo Scientific Halt Protease Inhibitor Single-Use Cocktails were tested against other commercially available tablet-format protease inhibitor cocktails under the same conditions. A 1X final concentration of each inhibitor was added. The single-use formulation resulted in ≥97% inhibition compared to ≥59% inhibition for the tablet.
Phosphatase inhibitors prevent the dephosphorylation of proteins, maintaining their phosphorylation status. These inhibitors contain sodium fluoride, sodium orthovanadate, sodium pyrophosphate, and β-glycerophosphate.
| | |
| Halt Phosphatase Inhibitor Cocktail | Pierce Phosphatase Inhibitor Tablet | |
| Format | Liquid | Tablet |
| Contains DMSO | Yes | No |
| Requires reconstitution | No (100X concentrate) | Yes |
| Downstream compatibility | Not compatible with 2D or IMAC | All |
| Available sizes | 24 x 100 uL 5 x 1 mL 1 mL 10 mL | Mini, 10 mL buffer per tablet |
| Documents | User Guide: Halt Phosphatase Inhibitor Cocktail | Product Sheet: Pierce Phosphatase Inhibitor Mini Tablets |
Broad-spectrum phosphatase inhibitors
Phosphatase inhibitors are available as a Halt Phosphatase Inhibitor Cocktail and Pierce Phosphatase Inhibitor Mini tablet formats. Both formats contain sodium fluoride, sodium orthovanadate, sodium pyrophosphate and beta-glycerophosphate inhibitors. Phosphatase inhibitors are compatible with cell lysis reagents.
The Halt Phosphatase Inhibitor Cocktail preserves the phosphorylation state of proteins by inhibiting serine/threonine phosphatases and tyrosine phosphatases during cell lysis or tissue protein extraction (Figure 3). It should be noted that the Halt phosphatase inhibitor cocktail interferes with 2D gel electrophoresis, and some immobilized metal chelate affinity chromatography methods but is compatible with the Pierce Phosphoprotein Enrichment Kit. Before performing incompatible procedures, either dialyze or desalt the sample to remove inhibitors from extracts.
The Pierce Phosphatase Inhibitor Mini tablets protect phosphorylation from serine/threonine phosphatases. The Pierce Phosphatase Inhibitor Mini tablet is suitable for use with any cell or tissue extract (Figure 4). To use, simply dissolve 1 phosphatase inhibitor mini tablet in 10 mL of solution.
Click image to enlarge
Figure 3. Preserved phosphorylation states of three proteins in cell lysate. HeLa cells were lysed in the absence (-) and presence (+) of a 1X concentration of the Thermo Scientific Halt Phosphatase Inhibitor Cocktail. Cell lysates were analyzed by Western blot for total and phosphorylated protein as indicated. Panel A: MEK and phosphorylated MEK (PMEK). Panel B: MAP Kinase and PMAP Kinase. Panel C: STAT3 and PSTAT3. The proteins are phosphorylated on serine, threonine/tyrosine and tyrosine, respectively.
Click image to enlarge
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 wothout inhibitor.
Protease and phosphatase cocktails preserve the native state of proteins and their post-translational modifications, helping ensure samples remain stable. These inhibitors contain cysteine protease inhibitors (E-64), serine protease inhibitors (Aprotinin, Leupeptin), and metalloprotease inhibitors (Bestatin, EDTA*) along with phosphatase inhibitors (sodium fluoride, sodium orthovanadate, sodium pyrophosphate, β-glycerophosphate).
| | | ||
| 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 | |
| Format | Liquid | Liquid, EDTA-free | Tablet | Tablet, 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 |
| Available sizes | 24 x 100 uL 5 x 1 mL 1 mL 10 mL | 24 x 100 uL 5 x 1 mL 1 mL 10 mL | Mini, 10 mL buffer per tablet | Mini, 10 mL buffer per tablet |
| Documents | Product Sheet: Halt Protease and Phosphatase Inhibitor Cocktail | Product Sheet: Pierce Protease and Phosphatase Inhibitor Mini Tablets | ||
*only in EDTA formulations
Combined protease and phosphatase inhibitors
Combined Protease and Phosphatase Inhibitors are available in cocktail and tablet formats. These broad-spectrum inhibitors protect against protein degradation while preserving protein phosphorylation simultaneously. They contain chemical compounds that inhibit serine-, cysteine-, and aspartic acid-proteases, and serine/threonine and tyrosine phosphatases, as well as aminopeptidases (Table 2). Metalloproteases are inhibited by the addition of EDTA, which is available in a separate vial in the cocktail format but included in the tablet format (Figure 5).
Table 2. Protease and phosphatase inhibitors included in the cocktail formulation and their targeted enzyme class.
Click image to enlarge
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.
Ordering information
See all: Protease and phosphatase inhibitors
Discover protein preparation tools
Protein sample preparation technical handbook
The Protein Preparation Handbook provides useful information on our broad selection of reagents and tools for protein extraction, clean-up, immunoprecipitation, and purification. Practical information, selection guides, and relevant data are included to help you improve your protein yield and downstream analysis.
Protein sample preparation eLearning course
This modular, animated, and narrated eLearning course was developed to provide a succinct, contextual summary of common laboratory methods and techniques required to achieve optimal results in protein assays and experiments. There are knowledge checks throughout the course to test what you have learned.
Related products and downstream applications
Hero banner dark gradient overlay
Stylesheet for Classic Wide Template adjustments
Hero banner dark gradient overlay
For Research Use Only. Not for use in diagnostic procedures.