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P8340 Sigma-Aldrich

Protease Inhibitor Cocktail

for use with mammalian cell and tissue extracts, DMSO solution

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Description

Specificity

Inhibits serine, cysteine, and acid proteases, and aminopeptidases.

Application

Protease Inhibitor Cocktail has been used-
• as a buffer component during sonication of GFP (green fluorescent protein)-huntingtin-transfected HEK 293 cells in GST (glutathione S-transferase) pull down assay
• as a component of lysis buffer
• as a component of radioimmunoprecipitation assay buffer (RIPA)

Packaging

1, 5 mL in serum bottle

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.

Components

AEBSF
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.

Safety & Documentation

Safety Information

WGK Germany 
WGK 2
Flash Point(F) 
188.6 °F - closed cup
Flash Point(C) 
87 °C - closed cup

Frequently Asked Questions

Is there anything in Product P8340, Protease Inhibitor Cocktail, that will cause downstream problems with RT-PCR?
While the product should not cause any downstream problems with RT-PCR, we would advise to clean up the reaction with either DMN10 (GenElute Direct mRNA Miniprep Kit) or T9424 (TRI Reagent®) for purification.
Can Product P8340, Protease Inhibitor Cocktail, be used with RIPA buffer and how much should be used?
Yes, this product can be used with RIPA buffer at 100 μl/ml concentration.
How much Product P8340, Protease Inhibitor Cocktail, should I use?
One mL of the cocktail solution is recommended for the inhibition of endogenous enzymes found in 100 mL of lysate from 20 g (wet weight) of bovine liver or 10 mL of cell lysate obtained from CHO cells at a cell density of 100 million (108) cells per mL. CHO cells were grown on DMEM with 10% FCS (heat inactivated).Note: Not all lysates contain the same levels of endogenous enzymes, and it may be necessary to adjust the volume of cocktail required.
What is the concentration of inhibitors in Product P8340, Protease Inhibitor Cocktail?
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.
DoesProduct P8340, Protease Inhibitor Cocktail, contain any chelators or EDTA?
This product does not contain any metal chelators.
Can Product P8340, Protease Inhibitor Cocktail, be used with Ezview Protein A and Anti FLAG M2 affinity gels.
Yes, this product can be used with both Ezview Protein A and Anti FLAG M2 affinity gels.
Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
How do I find price and availability?
There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote. USA customers: 1-800-325-3010 or view local office numbers.
What is the Department of Transportation shipping information for this product?
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.
My question is not addressed here, how can I contact Technical Service for assistance?
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Protocols & Articles

Articles

Protease Inhibitor Cocktails References

Our portfolio of protease inhibitor cocktails are frequently cited in peer-reviewed scientific literature, suggesting their superior quality and excellent track record. This page lists a sampling of ...
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Peer-Reviewed Papers
15

References

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