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.
P8340
Protease Inhibitor Cocktail
Protease Inhibitor Cocktail
1 of 4 reviewers received a sample product or took part in a promotion
DMSO solution, for the inhibition of serine, cysteine, acid proteases and aminopeptidases, for use with mammalian cell and tissue extracts, DMSO solution
Sinónimos:
Mezcla de inhibidor de proteasas
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Nombre del producto
Cóctel de inhibidores de proteasas, for use with mammalian cell and tissue extracts, DMSO solution
origen biológico
bovine lung
Nivel de calidad
Formulario
DMSO solution
solubilidad
water: soluble
temp. de almacenamiento
−20°C
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Categorías relacionadas
Descripción general
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.
Especificidad
Inhibe las proteasas de serina y cisteína, las proteasas ácidas y las aminopeptidasas.
Aplicación
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
Acciones bioquímicas o fisiológicas
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.
Características y beneficios
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
Componentes
AEBSF
Aprotinina
Bestatina
E-64
Leupeptina
Pepstatina A
Aprotinina
Bestatina
E-64
Leupeptina
Pepstatina A
Precaución
Este cóctel debe conservarse a -20 °C.
Nota de preparación
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.
Producto relacionado
Referencia del producto
Descripción
Precios
también adquirido normalmente con este producto
Referencia del producto
Descripción
Precios
Palabra de señalización
Warning
Frases de peligro
Consejos de prudencia
Clasificaciones de peligro
Eye Irrit. 2 - Skin Irrit. 2
Código de clase de almacenamiento
10 - Combustible liquids
Clase de riesgo para el agua (WGK)
WGK 2
Punto de inflamabilidad (°F)
188.6 °F - closed cup
Punto de inflamabilidad (°C)
87 °C - closed cup
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Certificados de análisis (COA)
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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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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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What is the working concentration if adding to the blood?
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The recommended amount to use is 1:100 (v:v) directly to the whole blood, but one has to take into consideration that this will disrupt the coagulation cascade. If it is a concern, the protease inhibitor cocktail should be added after separation of the blood components. In the serum use at the same concentration as for blood, 1:100 (v:v)
There are articles citing this product when using blood samples. 1-100 was used in both articles:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7346264/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2722770/Helpful?
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How long can this product be stored at -20C?
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This product is not assigned an expiration date. It is assigned a recommended retest date of approximately 4 years after the quality release date. A recommended retest date is the period of time during which the product is expected to remain within established stability specifications, provided that it has been stored under defined conditions. The retest date can be found in the lot specific Certificate of Analysis. After the Retest Date, product samples should be examined to ensure that the product is still in compliance with the established specifications. For more information, you may access the "Product Dating Information" document under "ADDITIONAL USEFUL DOCUMENTS ABOUT OUR PRODUCTS" at the bottom of the Quality Services page with this link: https://www.sigmaaldrich.com/US/en/life-science/quality-and-regulatory-management/quality-services.
Please see the link below to review a sample Certificate of Analysis:
https://www.sigmaaldrich.com/certificates/sapfs/PROD/sap/certificate_pdfs/COFA/Q14/P8340-1ML0000227406.pdfHelpful?
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Does Product P8340, Protease Inhibitor Cocktail, contain any chelators or EDTA?
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This product does not contain any metal chelators.
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Is there anything in Product P8340, Protease Inhibitor Cocktail, that will cause downstream problems with RT-PCR?
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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.
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