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
Cóctel de inhibidores de proteasas
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
Sinónimos:
Mezcla de inhibidor de proteasas
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Form:
DMSO solution
Solubility:
water: soluble
Storage temp.:
−20°C
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Permítanos ayudarleNombre del producto
Cóctel de inhibidores de proteasas, 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
Cóctel de inhibidores de proteasas para extractos de células y tejidos de mamíferos, en disolución DMSO. Contiene seis componentes individuales para inhibir las proteasas de serina, cisteína, las proteasas ácidas y las aminopeptidasas.
Application
Ideal para utilizar como componente del tampón durante el tratamiento con ultrasonidos, tampón de lisis y tampón para RIPA
Proporciona protección contra la actividad enzimática endógena en extractos celulares y de tejidos
Proporciona protección contra la actividad enzimática endógena en extractos celulares y de tejidos
Biochem/physiol Actions
Esta mezcla contiene componentes individuales, como la AEBSF a 104 mM, aprotinina a 80 μM, bestatina a 4 mM, E-64 a 1,4 mM, leupeptina a 2 mM y pepstatina A a 1,5 mM. Cada componente tiene propiedades inhibidoras específicas. La AEBSF y la aprotinina actúan para inhibir las proteasas de serina, como la tripsina, la quimiotripsina y la plasmina, entre otras. La bestatina inhibe las aminopeptidasas. La E-64 actúa contra las proteasas de cisteína. La leupeptina actúa contra las proteasas de serina y de cisteína. La pepstatina A inhibe las proteasas ácidas.
Inhibe las proteasas de serina y cisteína, las proteasas ácidas y las aminopeptidasas.
Features and Benefits
Inhibición de amplio espectro: inhibe múltiples clases de proteasas y aminopeptidasas
Conservación estable: se suministra como disolución en DMSO, lo que permite su conservación a largo plazo a -20°C
Rendimiento uniforme: disolución lista para usar
Conservación estable: se suministra como disolución en DMSO, lo que permite su conservación a largo plazo a -20°C
Rendimiento uniforme: disolución lista para usar
Preparation Note
Se recomienda 1 ml de esta disolución combinada para la inhibición de las enzimas endógenas que se encuentran en 100 ml de lisado procedente de 20 g de hígado bovino o 10 ml de lisado de células CHO. Este producto se suministra como una disolución en DMSO.
Other Notes
AEBSF
Aprotinina
Bestatina
E-64
Leupeptina
Pepstatina A
Aprotinina
Bestatina
E-64
Leupeptina
Pepstatina A
Disclaimer
Este cóctel debe conservarse a -20 °C.
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Este artículo | |||
|---|---|---|---|
| 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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Warning
hcodes
Hazard Classifications
Eye Irrit. 2 - Skin Irrit. 2
Clase de almacenamiento
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?
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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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