For saliva samples, P8340 is typically used at a 1:100 dilution (v:v), similar to its use in blood and tissue lysates. This means you would add 1 part of P8340 to 99 parts of saliva. However, the optimal concentration may need to be empirically determined for your specific application, as protease levels can vary between samples. There is no internal data on pre-diluting P8340 in water and storing it before use; the product is sold ready-to-use, and the best practice is to add it directly to your sample. If you wish to test pre-dilution and storage, you should validate this in your own experimental setup, as stability data for pre-diluted solutions is not available.
P8340
Cocktail di inibitori delle proteasi
DMSO solution, for the inhibition of serine, cysteine, acid proteases and aminopeptidases, for use with mammalian cell and tissue extracts, DMSO solution
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Sinonimo/i:
Mix di inibitori delle proteasi
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Informazioni su questo articolo
Form:
DMSO solution
Solubility:
water: soluble
Storage temp.:
−20°C
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Servizio Tecnico
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Permettici di aiutartiNome del prodotto
Cocktail di inibitori delle proteasi, for use with mammalian cell and tissue extracts, DMSO solution
biological source
bovine lung
Quality Segment
form
DMSO solution
input
mammalian sample(s)
solubility
water: soluble
storage temp.
−20°C
General description
Proteasi Cocktail di inibitori per estratti di cellule e tessuti di mammifero, fornito come soluzione in DMSO. Contiene sei singoli componenti per inibire la serin- e la cistein-proteasi, le proteasi acide e le amminopeptidasi.
Application
Ideale per l'uso come componente del tampone per sonicazione, tampone per lisi e tampone per RIPA
Fornisce protezione contro l'attività enzimatica endogena in estratti di cellule e tessuti
Fornisce protezione contro l'attività enzimatica endogena in estratti di cellule e tessuti
Biochem/physiol Actions
Inibisce le proteasi a serina, a cisteina, le proteasi acide e le amminopeptidasi.
Questa miscela contiene i singoli componenti, tra cui AEBSF 104 mM, aprotinina 80 μM, bestatina 4 mM, E-64 1,4 mM, leupeptina 2 mM e pepstatina A 1,5 mM. Ogni componente ha proprietà inibitrici specifiche. L'AEBSF e l'aprotinina sono in grado di inibire le proteasi a serina, tra cui la tripsina, la chimotripsina e la plasmina. La bestatina inibisce le amminopeptidasi. E-64 agisce contro le proteasi a cisteina. La leupeptina agisce contro le proteasi a serina e a cisteina La pepstatina A inibisce le proteasi acide.
Features and Benefits
Inibizione ad ampio spettro: inibisce più classi di proteasi e amminopeptidasi
Stabilità durante la conservazione: fornito come soluzione in DMSO, che ne consente la conservazione a lungo termine: a -20°C
Riproducibilità delle prestazioni: soluzione pronta all′uso
Stabilità durante la conservazione: fornito come soluzione in DMSO, che ne consente la conservazione a lungo termine: a -20°C
Riproducibilità delle prestazioni: soluzione pronta all′uso
Preparation Note
Si consiglia l′uso di 1 mL di questa soluzione cocktail per inibire gli enzimi endogeni presenti in 100 mL di lisato ottenuto da 20 g di fegato bovino o in 10 mL di lisato di cellule CHO. Questo prodotto viene fornito come soluzione in DMSO.
Other Notes
AEBSF
Aprotinina
Bestatina
E-64
Leupeptina
Pepstatina A
Aprotinina
Bestatina
E-64
Leupeptina
Pepstatina A
Disclaimer
Questo cocktail va conservato a -20 °C.
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Questo articolo | |||
|---|---|---|---|
| description for use with mammalian cell and tissue extracts, DMSO solution | description for use with bacterial cell extracts, lyophilized powder | description for general use, lyophilized powder | description - |
| form DMSO solution | form lyophilized powder | form lyophilized powder | form DMSO solution |
| biological source bovine lung | biological source - | biological source bovine lung | biological source - |
| Quality Level 300 | Quality Level 300 | Quality Level 300 | 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 |
| input mammalian sample(s) | input mammalian sample(s) | input general use sample(s) | input general use sample(s) |
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Parola di avvertenza
Warning
Codici di pericolo
Avvertenze di sicurezza
Hazard Classifications
Eye Irrit. 2 - Skin Irrit. 2
Classe di stoccaggio
10 - Combustible liquids
Punto di infiammabilità (°F)
188.6 °F - closed cup
Punto di infiammabilità (°C)
87 °C - closed cup
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What is the working concentration if adding to the whole saliva? Can I dilute it in water, store and then add to saliva?
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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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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.
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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?
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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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