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.
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P8340
Proteasehemmer-Cocktail
DMSO solution, for the inhibition of serine, cysteine, acid proteases and aminopeptidases, for use with mammalian cell and tissue extracts, DMSO solution
Protease Inhibitor Cocktail
Synonym(e):
Proteaseinhibitor-Mix
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Form:
DMSO solution
Solubility:
water: soluble
Storage temp.:
−20°C
Preise und Verfügbarkeit sind derzeit nicht verfügbar.
Technischer Dienst
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Unterstützung erhaltenProduktname
Proteasehemmer-Cocktail, for use with mammalian cell and tissue extracts, DMSO solution
biological source
bovine lung
Quality Segment
form
DMSO solution
solubility
water: soluble
storage temp.
−20°C
General description
Protease Inhibitor-Cocktail für Säugetierzell- und -gewebeextrakte, in DMSO-Lösung. Es enthält sechs einzelne Komponenten für die Hemmung von Serin-, Cystein- und Säureproteasen sowie Aminopeptidasen.
Application
Ideal zur Verwendung als Pufferkomponente bei der Beschallung, in Lysepuffer und RIPA-Puffer
Bietet Schutz vor endogener Enzymaktivität in Zell- und Gewebeextrakten
Bietet Schutz vor endogener Enzymaktivität in Zell- und Gewebeextrakten
Biochem/physiol Actions
Diese Mischung enthält individuelle Komponenten, einschließlich AEBSF zu 104 mM, Aprotinin zu 80 μM, Bestatin zu 4 mM, E-64 zu 1,4 mM, Leupeptin zu 2 mM und Pepstatin A zu 1,5 mM. Jede Komponente weist spezifische Hemmungseigenschaften auf. AEBSF und Aprotinin hemmen Serinproteasen, einschließlich unter anderem Trypsin, Chymotrypsin und Plasmin. Bestatin hemmt Aminpeptidasen. E-64 wirkt gegen Cysteinproteasen. Leupeptin wirkt gegen sowohl Serin- als auch Cysteinproteasen. Pepstatin A hemmt Säureproteasen.
Hemmt Serin-, Cystein-, und Säureproteasen sowie Aminopeptidasen.
Features and Benefits
Breitspektrumhemmung: hemmt mehrere Klassen von Proteasen und Aminopeptidasen
Stabile Lagerung: als Lösung in DMSO bereitgestellt, was eine Langzeitlagerung bei -20 °C ermöglicht
Gleichbleibende Leistung: gebrauchsfertige Lösung
Stabile Lagerung: als Lösung in DMSO bereitgestellt, was eine Langzeitlagerung bei -20 °C ermöglicht
Gleichbleibende Leistung: gebrauchsfertige Lösung
Preparation Note
Ein ml dieser Cocktaillösung wird für die Hemmung von endogenen Enzymen, die in 100 ml Lysat aus 20 g Rinderleber oder 10 ml Lysat aus CHO-Zellen vorkommen, empfohlen. Dieses Produkt wird als Lösung in DMSO bereitgestellt.
Other Notes
AEBSF
Aprotinin
Bestatin
E-64
Leupeptin
Pepstatin A
Aprotinin
Bestatin
E-64
Leupeptin
Pepstatin A
Disclaimer
Dieser Cocktail sollte bei -20 °C gelagert werden.
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Dieser Artikel | |||
|---|---|---|---|
| form DMSO solution | form DMSO solution | form DMSO solution | form DMSO solution |
| biological source bovine lung | biological source - | biological source - | biological source synthetic |
| Quality Level 300 | Quality Level 300 | Quality Level 300 | Quality Level 300 |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
| solubility water: soluble | solubility water: soluble | solubility water: soluble | solubility water: soluble |
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signalword
Warning
hcodes
Hazard Classifications
Eye Irrit. 2 - Skin Irrit. 2
Lagerklasse
10 - Combustible liquids
flash_point_f
188.6 °F - closed cup
flash_point_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?
1 answer-
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?
1 answer-
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
1 answer-
PMSF is a serine protease inhibitor. You can certainly add along with the cocktail if needed.
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