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P8340 Sigma

Protease Inhibitor Cocktail

for use with mammalian cell and tissue extracts, DMSO solution

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Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Components

AEBSF
Aprotinin
Bestatin
E-64
Leupeptin
Pepstatin A

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.

Caution

This cocktail should be stored at -20 °C.

Packaging

1, 5 mL in serum bottle

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.

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)

Price and Availability


Pharma QC

Amplified Detection

Biomedical Applications
Safety & Documentation

Safety Information

Symbol 
GHS07  GHS07
Signal word 
Warning
Hazard statements 
Precautionary statements 
WGK Germany 
2
Flash Point(F) 
188.6 °F
Flash Point(C) 
87 °C

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. This information can be viewed on our website by clicking on the following link:http://www.sigmaaldrich.com/sigma/general%20information/vol2%20issue3%20ez%20view.pdf
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.  This information can be found in P8340 datasheet by clicking on the following link: <rn:answer_xref style="TEXT-INDENT: 0in" title="P8340" target="_self" contents="http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Datasheet/5/p8340dat.Par.0001.File.tmp/p8340dat.pdf" answer_id="256" />
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 mMThis information can be viewed on our website by clicking on the following link:http://www.sigmaaldrich.com/sigma/general%20information/protinhibselectionguide.pdf
Does Product 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. This information can be viewed on our website by clicking on the following link:http://www.sigmaaldrich.com/sigma/general%20information/vol2%20issue3%20ez%20view.pdf
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. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
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, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
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Protocols & Articles

Articles

Protease Inhibitor Cocktails References

The Sigma-Aldrich portfolio of protease inhibitor cocktails shows an excellent track record in the peer-reviewed scientific literature, as witnessed by the many publications that cite our protease in...
Keywords: AGE, Adhesion, Aerobic, Affinity chromatography, Alzheimer Disease, Amplification, Angiogenesis, Anti-inflammatory agents, Antibiotics, Antimicrobials, Antitumor agents, Antivirals, Apoptosis, Atomic absorption spectroscopy, Autophagy, Biochemistry, Biological processes, Buffers, Cancer, Cardiovascular, Catalysis, Cell biology, Cell disruption, Cell proliferation, Cell signaling, Cellular processes, Central Nervous System, Chromatin immunoprecipitation, Chromatography, Clinical, Cloning, Cytotoxins, DNA replication, Degradations, Detergents, Diabetes, Diagnostic, Diseases, Dopamine agents, Drug discovery, Eliminations, Environmental, Enzyme-linked immunosorbent assay, Gas chromatography, Gastrointestinal, Gene expression, Genetic, Genetics, Gluconeogenesis, Glycosylations, Growth factors, Hormones, Hydration reaction, Hydroxylations, Immobilization, Immunology, Immunoprecipitation, Inflammation, Ligands, Lipid Metabolism, Lipid peroxidation, Mass spectrometry, Medicinal chemistry, Metabolism, Metabolites, Methylations, Microarray Analysis, Microbiology, Microscopy, Molecular biology, Morphogenesis, Neurology, Neuroscience, Neurotransmission, Neurotransmitters, Normal-phase chromatography, Nutrition, Obesity, Oncology, PAGE, Parkinson Disease, Peroxidations, Pharmaceutical, Phosphorylations, Photosynthesis, Physiological Processes, Polymerase chain reaction, Polymorphisms, Proteomics, Purification, Recombination, Reductions, Reproduction, Respiratory, Reversible addition-fragmentation chain transfer polymerizations, Sample preparations, Scanning electron microscopy, Schizophrenia, Separation, Sequencing, Sonication, Transcription, Transduction, Veterinary, Vitamins, Western blot, transformation

Peer-Reviewed Papers
15

References

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