Pierce protease inhibitor cocktails and phosphatase inhibitor formats

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.

Cell Lysis Buffers  Cell Fractionation Kits

Broad-spectrum protease inhibitor cocktails

 LiquidLiquid, EDTA-freeTabletTablet, EDTA-free
 
 Halt Protease Inhibitor CocktailHalt Protease Inhibitor Cocktail, EDTA-free
Contains EDTAYes, but in a separate vialNoYesNo
Contains DMSOYesYesNoNo
Requires reconstitutionNo (100X concentrate)No (100X concentrate)YesYes
Downstream compatibilityNot compatible with 2D or IMACAllNot compatible with 2D or IMACAll
Included inhibitorsAEBSF, 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

Graph demonstrating effective broad-spectrum protease inhibition by Pierce Protease Inhibitor Cocktail Tablets in various lysates

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

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

 LiquidTablet
 
 Halt Phosphatase Inhibitor CocktailPierce Phosphatase Inhibitor Tablet
Contains DMSOYesNo
Requires reconstitutionNo (100X concentrate)Yes
Downstream compatibilityNot compatible with 2D or IMACAll
Included inhibitorsSodium 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

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

Inhibition of phosphatase activity by Pierce Phosphatase inhibitors on Liver and Spleen protein extracts
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

 LiquidLiquid, EDTA-freeTabletTablet, EDTA-free
 Halt Protease and Phosphatase Inhibitor CocktailHalt Protease and Phosphatase Inhibitor Cocktail, EDTA-freePierce Protease and Phosphatase Inhibitor TabletsPierce Protease and Phosphatase Inhibitor Tablets, EDTA-free
Contains EDTAYes, but in a separate vialNoYesNo
Included inhibitorsAprotinin, bestatin, E-64, leupeptin, sodium fluoride, sodium orthovanadate, sodium pyrophosphate, β-glycerophosphate, EDTA*
Contains DMSOYesYesNoNo
Requires reconstitutionNo (100X concentrate)No (100X concentrate)YesYes
Downstream compatibilityNot compatible with 2D or IMACAllNot compatible with 2D or IMACAll
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.

Combined Pierce protease and phosphatase inhibitor cocktails effectiveness on protease and phosphatase activity in pancreatic tissue
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.

Broad-spectrum protease inhibitor cocktails

 LiquidLiquid, EDTA-freeTabletTablet, EDTA-free
 
 Halt Protease Inhibitor CocktailHalt Protease Inhibitor Cocktail, EDTA-free
Contains EDTAYes, but in a separate vialNoYesNo
Contains DMSOYesYesNoNo
Requires reconstitutionNo (100X concentrate)No (100X concentrate)YesYes
Downstream compatibilityNot compatible with 2D or IMACAllNot compatible with 2D or IMACAll
Included inhibitorsAEBSF, 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

Graph demonstrating effective broad-spectrum protease inhibition by Pierce Protease Inhibitor Cocktail Tablets in various lysates

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

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

 LiquidTablet
 
 Halt Phosphatase Inhibitor CocktailPierce Phosphatase Inhibitor Tablet
Contains DMSOYesNo
Requires reconstitutionNo (100X concentrate)Yes
Downstream compatibilityNot compatible with 2D or IMACAll
Included inhibitorsSodium 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

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

Inhibition of phosphatase activity by Pierce Phosphatase inhibitors on Liver and Spleen protein extracts
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

 LiquidLiquid, EDTA-freeTabletTablet, EDTA-free
 Halt Protease and Phosphatase Inhibitor CocktailHalt Protease and Phosphatase Inhibitor Cocktail, EDTA-freePierce Protease and Phosphatase Inhibitor TabletsPierce Protease and Phosphatase Inhibitor Tablets, EDTA-free
Contains EDTAYes, but in a separate vialNoYesNo
Included inhibitorsAprotinin, bestatin, E-64, leupeptin, sodium fluoride, sodium orthovanadate, sodium pyrophosphate, β-glycerophosphate, EDTA*
Contains DMSOYesYesNoNo
Requires reconstitutionNo (100X concentrate)No (100X concentrate)YesYes
Downstream compatibilityNot compatible with 2D or IMACAllNot compatible with 2D or IMACAll
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.

Combined Pierce protease and phosphatase inhibitor cocktails effectiveness on protease and phosphatase activity in pancreatic tissue
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.

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