The actual volume of the solution is determined by the lot-specific activity, which can be found in the Certificate of Analysis. For this product, there are 5,000 units available. If the lot has a specific activity of 335 U/?L, the volume will be approximately 14.9 ?L for 5,000 units. It is recommended to dilute the product in 20 mM Tris-HCl (pH 8.0), 2 mM MgCl?, and 20 mM NaCl.
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5 KU
CHF 236.00
25 KU
CHF 807.00
Informazioni su questo articolo
Numero CAS:
UNSPSC Code:
12352204
NACRES:
NA.54
MDL number:
Assay:
≥90% (SDS-PAGE)
Biological source:
Serratia marcescens
Recombinant:
expressed in E. coli
Concentration:
≥250 units/μL
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Servizio Tecnico
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Permettici di aiutartibiological source
Serratia marcescens
recombinant
expressed in E. coli
assay
≥90% (SDS-PAGE)
form
buffered aqueous glycerol solution
mol wt
30 kDa
concentration
≥250 units/μL
application(s)
research use
foreign activity
protease, essentially free
shipped in
wet ice
storage temp.
−20°C
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Questo articolo | |||
|---|---|---|---|
| assay ≥90% (SDS-PAGE) | assay >99% (SDS-PAGE) | assay >99% (SDS-PAGE) | assay >90% (SDS-PAGE) |
| biological source Serratia marcescens | biological source Serratia marcescens | biological source Serratia marcescens | biological source Serratia marcescens |
| form buffered aqueous glycerol solution | form buffered aqueous glycerol solution | form buffered aqueous glycerol solution | form buffered aqueous glycerol solution |
| application(s) research use | application(s) research use | application(s) research use | application(s) research use |
| concentration ≥250 units/μL | concentration 25-29 units/μL | concentration ≥250 units/μL | concentration ≥250 units/μL |
| recombinant expressed in E. coli | recombinant expressed in E. coli | recombinant expressed in E. coli | recombinant expressed in E. coli |
General description
Benzonase® è un'endonucleasi geneticamente modificata, altamente efficiente, estratta da Serratia marcescens[1]. Questa proteina dimerica caratterizzata da due ponti disolfuro essenziali [2] è in grado di attaccare e degradare tutte le forme di DNA e RNA (a singolo o doppio filamento, lineare e circolare) ed è efficace in una vasta gamma di condizioni operative. La nucleasi Benzonase® è in grado di rimuovere gli acidi nucleici e migliorare la purezza e la qualità dei campioni di proteine.
Application
Utilizzato per rimuovere gli acidi nucleici dai campioni di proteine.
La nucleasi Benzonase® è stata impiegata: come componente in un tampone di lisi C ghiacciato per la digestione di DNA e RNA per facilitare il rilascio completo di tutte le proteine nucleari nella fase di immunoprecipitazione per liberare i complessi proteici dal nucleoplasma e dalla cromatinacome supplemento nel tampone RIPA per il frazionamento delle cellule SHSY5Y per l'immunoprecipitazione per rimuovere gli acidi nucleici residui dalle radici aortiche con il metodo della decellularizzazione
Biochem/physiol Actions
Digerisce DNA ed RNA nativi o denaturati con calore.
La nucleasi Benzonase® può digerire completamente gli acidi nucleici a oligonucleotidi 5′-monofosfato lunghi 3-5 basi, caratteristica che la rende ideale per la rimozione degli acidi nucleici dalle proteine ricombinanti e per le applicazioni che richiedono la digestione completa degli acidi nucleici. Oltre a ridurre la viscosità degli estratti proteici e a evitare la formazione di aggregati di cellule, il pretrattamento dei campioni di proteine con la nucleasi Benzonase® può migliorare significativamente la risoluzione all'elettroforesi 2D su gel eliminando tutti gli acidi nucleici legati. Questo enzima versatile è in grado di digerire il DNA e l'RNA sia nella forma nativa che in quella denaturata al calore, al suo pH ottimale per l'attività enzimatica di 8,0-9,2. La nucleasi Benzonase® è efficace nel rimuovere il DNA dell'ospite nei campioni di microbioma. In molti casi, i campioni di microbioma (salivari o cutanei) contengono un'elevata percentuale di DNA dell'ospite che interferisce con i risultati delle applicazioni a valle. I nostri esperti hanno dimostrato che la riduzione del DNA dell'ospite abbassa i costi del sequenziamento mentre arricchisce e migliora i dati. I dati sperimentali sono presentati nell'articolo tecnico - La nucleasi Benzonase® per i flussi operativi con il microbioma
Features and Benefits
- Deplezione del DNA dell'ospite in campioni di microbioma.
- Digestione efficace degli acidi nucleici in diversi tipi di sequenze operative
- Riduzione della viscosità durante l'estrazione delle proteine.
Physical form
Soluzione in glicerolo al 50% contenente Tris HCl 20 mM, pH 8,0, MgCl22 mM e NaCl 20 mM.
Other Notes
Un'unità rappresenta la quantità di enzima in grado di digerire il DNA di sperma di salmone sonicato a oligonucleotidi solubili in acido equivalenti a ΔA260 pari a 1,0 in 30 minuti a pH 8,0 e a 37 °C (volume di reazione 2,625 mL)
Legal Information
La nucleasi Benzonase® è fornita da Merck KGaA, Darmstadt, Germania e/o dalle sue affiliate.
Benzonase is a registered trademark of Merck KGaA, Darmstadt, Germany
Classe di stoccaggio
10 - Combustible liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves
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Numero articolo commerciale globale
| SKU | GTIN |
|---|---|
| E1014-5KU | 04061835572922 |
| E1014-25KU | 04061835572915 |
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hi, I received E1014-5KU product. I couldn't find what solution should I dilute to get a concentration of 25U/ml. Also, the volume of the solution.
1 answer-
Helpful?
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Do you sell high salt lysis buffers that are compatible with benzonase? Do you have any written protocols for use of benzonase with high salt buffers?
1 answer-
We do not have a branded "high-salt lysis buffer" off the shelf that is compatible with benzonase. However, we offer a specialized Benzonase® Salt Tolerant Endonuclease which is active at higher salt concentration, up to1M NaCl .
https://www.sigmaaldrich.com/US/en/product/mm/e5014Helpful?
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I am doing a protein purification and would like to know if magnesium chloride is required in the lysis buffer when using the Benzonase. Also, would the activity of benzonase be affected when added to a lysis buffer containing EDTA
1 answer-
A concentration of 1-2 mM of magnesium is required for the activity of this product. An EDTA concentration of 1 mM partially inhibits the activity of this product.
Helpful?
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Hi, what is the ratio of E1014 : DNA and E1014 : RNA for an efficient acid nucleic removal?
1 answer-
For efficient nucleic acid removal using Benzonase® (E1014), the recommended ratio depends on the concentration of DNA or RNA in the sample. A typical starting point is 25 U/mL of Benzonase® for reducing nucleic acid content in lysates, which corresponds to approximately 1 unit of enzyme per 37 µg of DNA under optimal conditions. For lower concentrations of DNA or RNA, as little as 9 U/mL may achieve 99% removal, but higher concentrations, e.g., 90 U/mL, ensure faster and more complete degradation. Optimal activity requires 1–2 mM magnesium ions, a pH of 8.0–9.2, and incubation at 37°C. Adjustments may be needed for sample-specific factors such as buffer composition and nucleic acid load.
Helpful?
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How many ul does it contain? What kind of Unit is KU?
1 answer-
The unit activity of this product is lot-specific and reported in the product Certificate of Analysis. The minimum activity is 250 units per microliter. The KU value represents 1000 units. For example, a 20 KU package size represents 20,000 units. Please see the link below to review a sample or lot-specific Certificate:
https://www.sigmaaldrich.com/product/sigma/e1014#product-documentationHelpful?
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Hi, for protein purification from E. coli cells, how much is the working concentration range for Benzonase nuclease (≥250 units/μL)? Thank you
1 answer-
The recommended starting concentration for Benzonase nuclease is 25 units per milliliter of cell lysate.
Helpful?
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Hello, I plan to do proteomic for protein. When I add RIPA buffer to isolate the protein, I need to remove the DNA and RNA inside. When should I add E1014? Together with RIPA or after it?
1 answer-
Concentrations greater than 1 mM EDTA will inhibit Benzonase activity, and RIPA buffer recipes typically contain EDTA at higher concentrations than 1 mM. Therefore, Benzonase should be used after removing EDTA from the lysed sample or consider using a different lysis solution that does not include EDTA in the formulation.
Helpful?
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What is the storage temperature range for E1014? There is only a specified storage temperature, and not a range.
1 answer-
This product is stored at freezer temperature, which is typically -20°C. An excepted range is -15 - -20°C.
Helpful?
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