Common protease inhibitors used in chromatin immunoprecipitation assays include PMSF, EDTA, Leupeptin, Aprotinin, and Pepstatin A. This product contains multiple protease inhibitors to provide broad-spectrum inhibition, making it suitable for downstream ChIP experiments.
P8340
Protease Inhibitor Cocktail
DMSO solution, for the inhibition of serine, cysteine, acid proteases and aminopeptidases, for use with mammalian cell and tissue extracts, DMSO solution
Protease Inhibitor Cocktail
Synonym(s):
Protease Inhibitor Mix
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About This Item
Form:
DMSO solution
Solubility:
water: soluble
Storage temp.:
−20°C
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Product Name
Protease Inhibitor Cocktail, for use with mammalian cell and tissue extracts, DMSO solution
biological source
bovine lung
Quality Level
form
DMSO solution
solubility
water: soluble
storage temp.
−20°C
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Related Categories
1 of 4
This Item | P9599 | P8215 | P8849 |
|---|---|---|---|
| form DMSO solution | form DMSO solution | form DMSO solution | form DMSO solution |
| Quality Level 300 | Quality Level 300 | Quality Level 300 | Quality Level 300 |
| biological source bovine lung | biological source - | biological source - | biological source synthetic |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
General description
Protease Inhibitor Cocktail for mammalian cell and tissue extracts, in DMSO solution. It contains six individual components for inhibiting serine, cysteine, and acid proteases, and aminopeptidases.
Application
Ideal for use as a buffer component during sonication, lysis buffer, and RIPA buffer
Provides protection against endogenous enzyme activity in cell and tissue extracts
Provides protection against endogenous enzyme activity in cell and tissue extracts
Biochem/physiol Actions
Inhibits serine, cysteine, and acid proteases, and aminopeptidases.
This mixture contains individual components, including AEBSF at 104 mM, Aprotinin at 80 μM, Bestatin at 4 mM, E-64 at 1.4 mM, Leupeptin at 2 mM and Pepstatin A at 1.5 mM. Each component has specific inhibitory properties. AEBSF and Aprotinin act to inhibit serine proteases, including trypsin, chymotrypsin, and plasmin amongst others. Bestatin inhibits aminpeptidases. E-64 acts against cystein proteases. Leupeptin acts against both serine and cystein proteases. Pepstatin A inhibits acid proteases.
Features and Benefits
Broad-spectrum inhibition: inhibits multiple classes of proteases and aminopeptidases
Stable storage: supplied as a solution in DMSO, allowing for long-term storage at -20°C
Consistent performance: ready-to-use solution
Stable storage: supplied as a solution in DMSO, allowing for long-term storage at -20°C
Consistent performance: ready-to-use solution
Preparation Note
One mL of this cocktail solution is recommended for the inhibition of endogenous enzymes found in 100 mL of lysate from 20 g of bovine liver or 10 mL of lysate from CHO cells. This product is supplied as a solution in DMSO.
Other Notes
AEBSF
Aprotinin
Bestatin
E-64
Leupeptin
Pepstatin A
Aprotinin
Bestatin
E-64
Leupeptin
Pepstatin A
Disclaimer
This cocktail should be stored at -20 °C.
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Description
Pricing
Signal Word
Warning
Hazard Statements
Precautionary Statements
Hazard Classifications
Eye Irrit. 2 - Skin Irrit. 2
Storage Class Code
10 - Combustible liquids
WGK
WGK 2
Flash Point(F)
188.6 °F - closed cup
Flash Point(C)
87 °C - closed cup
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Select different protease inhibitor types based on your needs to prevent protein degradation during isolation and characterization and safeguard proteins in sample prep.
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Can I add the the protease inhibitor to RIPA as 100ul of inhibitor and 10ml RIPA buffer, aliquots and store them at -20C ?
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Is this product suitable for downstream ChIP experiments?
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Will the protease inhibitors in this product remain stable/functional through repeated freeze/thaw cycles? If not, is there a recommended limit to the number of freeze/thaw cycles? Should this product be aliquoted?
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We have not studied P8340 product for stability in repeated freeze/thaw cycles. Internally, we have made a 25 mg/mL stock solution; by dissolving 500 mg in 20 ml of water; this stock is around 100 mM (100x); We then aliquot 1mL and freeze once at -20C.
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Can this product be used for IL-6 detection using ELISA in rat tissue Homogenate? Can it be used along with PMSF without creating any problem in the assay?
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This product can be used in rat tissue homogenate. It already contains serine and cysteine protease inhibitors. Note that PMSF has a short half-life in aqueous solutions, specifically 55 minutes at 25°C and pH 7.5.
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How long is the protease inhibitor cocktail stable at room temperature?
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This product ships without wet or dry ice and is stable for up to two weeks at ambient temperatures. The long-term stability of this material at room temperature has not been determined. The recommended long-term storage temperature is -20°C.
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Can we dilute it in water? The experiment I plan to do will require 100ul of Protease inhibitor per vial, and I have more than 50 samples to run.
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This product is ready to use and should be diluted into the lysis buffer.
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Is the protease cocktail P8340 supplied as a 100X solution or a 1X solution?
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This cocktail is essentially a 100X preparation. One milliliter of this solution is sufficient for the inhibition of endogenous enzymes found in 100 mL of lysate. Please see the link below to review this and additional information available in the product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/107/629/p8340dat-ms.pdfHelpful?
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What is the stock concentration of the Protease Inhibitor Cocktail P8340? Is it 1x or 100x?
1 answer-
The concentration of inhibitors in this product is the following:AEBSF, 104 mMAprotinin, 0.085 mMBestatin, 4 mME-64, 1.4 mMLeupeptin, 2 mMPepstain A, 1.5 mM.
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Do we need to add PMSF as well along with this cocktail? I intend to use this for RIPA buffer
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PMSF is a serine protease inhibitor. You can certainly add along with the cocktail if needed.
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What is the working concentration if adding to the blood?
1 answer-
The recommended amount to use is 1:100 (v:v) directly to the whole blood, but one has to take into consideration that this will disrupt the coagulation cascade. If it is a concern, the protease inhibitor cocktail should be added after separation of the blood components. In the serum use at the same concentration as for blood, 1:100 (v:v)
There are articles citing this product when using blood samples. 1-100 was used in both articles:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7346264/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2722770/Helpful?
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