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.
P8340
Protease Inhibitor Cocktail
Protease Inhibitor Cocktail
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DMSO solution, for the inhibition of serine, cysteine, acid proteases and aminopeptidases, for use with mammalian cell and tissue extracts, DMSO solution
Synonym(s):
Protease Inhibitor Mix
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About This Item
form:
DMSO solution
storage temp.:
−20°C
Pricing and availability is not currently available.
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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
storage temp.
−20°C
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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.
Specificity
Inhibits 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
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
Components
AEBSF
Aprotinin
Bestatin
E-64
Leupeptin
Pepstatin A
Aprotinin
Bestatin
E-64
Leupeptin
Pepstatin A
Caution
This cocktail should be stored at -20 °C.
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.
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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 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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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?
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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?
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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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How long can this product be stored at -20C?
1 answer-
This product is not assigned an expiration date. It is assigned a recommended retest date of approximately 4 years after the quality release date. A recommended retest date is the period of time during which the product is expected to remain within established stability specifications, provided that it has been stored under defined conditions. The retest date can be found in the lot specific Certificate of Analysis. After the Retest Date, product samples should be examined to ensure that the product is still in compliance with the established specifications. For more information, you may access the "Product Dating Information" document under "ADDITIONAL USEFUL DOCUMENTS ABOUT OUR PRODUCTS" at the bottom of the Quality Services page with this link: https://www.sigmaaldrich.com/US/en/life-science/quality-and-regulatory-management/quality-services.
Please see the link below to review a sample Certificate of Analysis:
https://www.sigmaaldrich.com/certificates/sapfs/PROD/sap/certificate_pdfs/COFA/Q14/P8340-1ML0000227406.pdfHelpful?
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What is the Department of Transportation shipping information for this product?
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Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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Can Product P8340, Protease Inhibitor Cocktail, be used with Ezview Protein A and Anti FLAG M2 affinity gels.
1 answer-
Yes, this product can be used with both Ezview Protein A and Anti FLAG M2 affinity gels.
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