For use with blood, add the stock solution at a 1:10 dilution, corresponding to 100 µL of reconstituted cocktail per 1 mL of blood, to achieve a 1X final concentration. The optimal concentration may vary depending on the specific application. If there is an anticipated delay in processing whole blood samples, the addition of protease inhibitors may help minimize protein degradation.
P2714
Protease Inhibitor Cocktail
lyophilized powder, for the inhibition of serine, cysteine and metallo-proteases, for general use, lyophilized powder
Synonym(s):
Protease Inhibitor Cocktail from bovine lung, Protease Inhibitor Mix, Protease inhibitor
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| Size/SKU | Availability | Price |
|---|---|---|
1 btl | Check Cart for Availability | $89.60 |
About This Item
UNSPSC Code:
12352200
NACRES:
NA.77
Form:
lyophilized powder
Solubility:
water: soluble
Storage temp.:
−20°C
Product Name
Protease Inhibitor Cocktail powder, for general use, lyophilized powder
biological source
bovine lung
Quality Segment
form
lyophilized powder
input
general use sample(s)
solubility
water: soluble
storage temp.
−20°C
General description
The protease inhibitor cocktail has been optimized and tested for general use. It consists of water-soluble protease inhibitors that effectively inhibit serine, cysteine, and metalloproteases. This product is supplied as a lyophilized powder.
Application
Protease Inhibitor Cocktail powder has been used as a component:
- in radioimmunoprecipitation assay buffer to lyse the avian crystalline lens samples for western blot analysis
- in phosphate buffer solution (PBS) for grinding recombinant protein expressed leaves to purify virus-like particles (VLPs)
- in PBS to dissolve lyophilized skin secretion sample of A. loloensis for peptide purification
- in 0.1 M NaCl solution to wash and collect skin secretions of frog
Biochem/physiol Actions
Inhibits serine, cysteine, and metallo-proteases.
The components of this mixture, such as AEBSF and Aprotinin act to inhibit serine proteases, including trypsin, chymotrypsin, and plasmin amongst others. Bestatin inhibits aminpeptidases. E-64 acts against cystein proteases. EDTA is an inhibitor of metalloproteases. Leupeptin acts against both serine and cystein proteases.
Preparation Note
One bottle makes 100 mL of cocktail, which are sufficient to the inhibition of proteases present in approximately 20 gram of cell extract.
Not all extracts contain the same levels of endogenous enzymes, and it may be necessary to adjust the volume of cocktail required.
Not all extracts contain the same levels of endogenous enzymes, and it may be necessary to adjust the volume of cocktail required.
This product is supplied as a lyophilized powder. One bottle prepares 100 mL of a cocktail solution.
Other Notes
For R&D use only. Not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices.
This mixture contains individual components, including:
AEBSF at 2 mM
Aprotinin at 0.3 μM
Bestatin at 116 μ
E-64 at 14 μM
EDTA at 1 mM
Leupeptin at 1 μM
Each component has specific inhibitory properties.
AEBSF at 2 mM
Aprotinin at 0.3 μM
Bestatin at 116 μ
E-64 at 14 μM
EDTA at 1 mM
Leupeptin at 1 μM
Each component has specific inhibitory properties.
Disclaimer
The lyophilized powder is stable for 4 years when stored at -20°C.
1 of 1
This Item | |||
|---|---|---|---|
| description for general use, lyophilized powder | description for use with bacterial cell extracts, lyophilized powder | description for general use, lyophilized powder | description for use in tissue culture media, DMSO solution |
| form lyophilized powder | form lyophilized powder | form lyophilized powder | form DMSO solution |
| biological source bovine lung | biological source - | biological source - | biological source - |
| Quality Level 300 | Quality Level 300 | Quality Level 200 | Quality Level 200 |
| solubility water: soluble | solubility water: soluble | solubility water: soluble | solubility water: soluble |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
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signalword
Warning
hcodes
Hazard Classifications
Eye Irrit. 2 - Skin Irrit. 2
Storage Class
11 - Combustible Solids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
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How much of the reconstituted cocktail should I add to 1mL of blood?
1 answer-
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Can I dissolve this product in DMSO (for increased shelf life of the aliquotes at -80degC) rather than water?
1 answer-
This product is water-soluble. The solubility in DMSO is have not been investigated. However, DMSO at concentrations above 1% can destabilize the protein structure.
The product is packaged in a 10 mL serum vial with a crimp cap. Dissolve the contents of the vial in 10 mL of working buffer, then transfer to another container for dilution to 100 mL. Store the solution in working aliquots for at least 1 month -20 C, to avoid repeated freeze/thaw cycles. Please see the link below to review additional information available in the product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/212/061/p2714dat-ms.pdfThere are several Protease Inhibitor Cocktail formulations that are prepared in DMSO that may be of interest. See the links available below to review these options.
https://www.sigmaaldrich.com/US/en/product/sigma/p1860
https://www.sigmaaldrich.com/US/en/product/sigma/p8215
https://www.sigmaaldrich.com/US/en/product/sigma/p8340Helpful?
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What reagent and volume do I reconstitute this vial?
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One bottle prepares 100 mL of cocktail solution. The product is packaged in a 10 mL serum vial with a crimp cap. Dissolve the contents of the vial in 10 mL of water or of any working lysis buffer, then transfer to another container for dilution to 100 mL.
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In un estratto vegatale di 1 mL quanto microL devo aggiungere di cocktail?
1 answer-
Not all extracts contain the same levels of endogenous enzymes, so it is necessary to empirically determine the required volume of cocktail. For reference, one mL of the P2714 cocktail solution is recommended for the inhibition of endogenous enzymes equivalent to 1 mg of USP pancreatin. One bottle is recommended for the inhibition of proteases present in a maximum of 20 g of cell extract.
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what is the concentration of P2174 per skin/tissue piece?
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We do not measure the concentration of P2714 per skin/tissue piece. Instead, the measured components are: AEBSF at 2 mM, Aprotinin at 0.3 μM, Bestatin at 116 μM, E-64 at 14 μM, Leupeptin at 1 μM and EDTA at 1 mM. The amount in each vial is 89 mg. This material is very hygroscopic. It is recommended to prepare the entire package rather than attempt to weigh out specific amounts.
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How much PIC do I need to use in salivary samples for Western Blot?
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One bottle of the lyophilized powder makes 100 mL of the cocktail. For salivary samples, a 1:500 dilution factor has been known to work well.
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Hi Dose this product P2714, include EDTA? what proteases do it inhibit? Thanks, Camilla
1 answer-
P2714 includes EDTA at 1 mM. The components of this mixture, such as AEBSF and aprotinin, act to inhibit serine proteases, including trypsin, chymotrypsin, and plasmin among others. Bestatin inhibits aminpeptidases. E-64 acts against cystein proteases. EDTA is an inhibitor of metalloproteases. Leupeptin acts against both serine and cystein proteases.
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Wie löse ich das Pulver in 100 ml aqua dest???? Wie lagere ich das dann?
1 answer-
This product is packaged in a 10 mL serum vial with a crimp cap. Dissolve the contents of the vial in 10 mL of water or working buffer, then transfer to another container for dilution to 100 mL. Store the solution in working aliquots for at least 1 month -20 C, to avoid repeated freeze/thaw cycles. See the link below to review the product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/212/061/p2714dat-ms.pdfHelpful?
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I intend on taking 1mL samples of blood to give serum and test hormone levels in these samples. Should I add the solubilised protease inhibitor cocktail (p2714) directly to the blood or after centrifugation (which could be a few hours later)?
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A short delay may be negligible, however, it would not be detrimental to add protease inhibitors to whole blood samples if an extended delay is anticipated. This would ensure the best results. Please see the link below to review a publication of a similar nature:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6817059/Helpful?
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How much powder is in each bottle?
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The amount in each vial is 89 mg. This material is very hygroscopic. It is recommended to prepare the entire package rather than attempt to weigh out specific amounts.
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