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.
E1014
Benzonase® Nuklease
≥250 units/μL, ≥90% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution
Benzonase® Nuclease
Synonym(e):
Endonuclease aus Serratia marcescens
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5 KU
CHF 236.00
25 KU
CHF 807.00
Über diesen Artikel
CAS-Nummer:
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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Unterstützung erhaltenbiological 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
Quality Level
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Verwandte Kategorien
1 of 4
Dieser Artikel | |||
|---|---|---|---|
| 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®-Nuklease ist eine besonders effiziente und gentechnisch veränderte Endonuklease aus Serratia marcescens[1]. Dieses dimere Protein mit zwei essenziellen Disulfidbindungen [2] kann alle Formen von DNA und RNA (einzelsträngig, doppelsträngig, linear und zirkulär) unter einem breiten Spektrum von Arbeitsbedingungen angreifen und abbauen. Benzonase®-Nuklease kann aufgrund ihrer Fähigkeit, Nukleinsäuren zu entfernen, die Reinheit und Qualität von Proteinproben verbessern.
Application
Entfernung von Nukleinsäure in Proteinproben
Benzonase-®Nuklease wird wie folgt verwendet: als Komponente im eiskalten Lysepuffer C zum Verdau von DNA und RNA, um die vollständige Freisetzung aller nukleären Proteine zu erleichtern im Immunpräzipitationsschritt, um Proteinkomplexe aus Nukleoplasma und Chromatin freizusetzen als Supplement in RIPA zur Fraktionierung von SHSY5Y-Zellen für die Immunpräzipitation zur Entfernung von Nukleinsäureresten aus den Aortenwurzeln beim Dezellularisierungsverfahren
Biochem/physiol Actions
Benzonase®-Nuklease kann Nukleinsäuren vollständig zu Oligonukleotiden mit endständigem 5′-Monophosphat und einer Länge von 3 bis 5 Basen abbauen. Dies macht sie zu einem idealen Tool für das Entfernen von Nukleinsäuren aus rekombinanten Proteinen und für Anwendungen, die einen vollständigen Verdau von Nukleinsäuren erfordern. Neben dem Herabsetzen der Viskosität in Proteinextrakten und dem Verhindern des Verklumpens von Zellen kann die Vorbehandlung von Proteinproben mit Benzonase®-Nuklease ihre Auflösung bei der 2D-Gelelektrophorese signifikant verbessern, indem sie alle gebundenen Nukleinsäuren entfernt. Dieses vielseitige Enzym kann sowohl native als auch hitzedenaturierte DNA und RNA abbauen. Der optimale pH-Wert für seine Enzymaktivität liegt bei 8,0 bis 9,2. Benzonase® Nuklease ist wirksam beim Entfernen von Wirts-DNA aus Mikrobiomproben. In vielen Fällen weisen Mikrobiomproben (wie Speichel oder Haut) einen hohen Anteil Wirts-DNA auf, die Downstream-Ergebnisse beeinträchtigt. Unsere Experten zeigen, dass die Verringerung der Wirts-DNA die Kosten der Sequenzierung senkt und gleichzeitig die Daten verbessert. Experimentelle Daten werden in diesem Fachbeitrag gezeigt: Benzonase® Nuclease for Microbiome Workflows
Verdaut native oder hitzedenaturierte DNA und RNA.
Features and Benefits
- Depletion der Wirts-DNA in Mikrobiom-Proben.
- Effektiver Nukleinsäureverdau in verschiedenen Arbeitsabläufen.
- Reduktion der Viskosität bei der Proteinextraktion.
Physical form
Lösung in 50% Glycerin, enthält 20 mM Tris HCl, pH 8.0, 2 mM MgCl2, and 20 mM NaCl.
Other Notes
Eine Einheit verdaut mit Ultraschall behandelte Lachssperma-DNA in säurelösliche Oligonukleotide entsprechend einem ΔA260 von 1,0 in 30 min bei einem pH-Wert von 8,0 bei 37°C (Reaktionsvolumen 2,625 ml).
Legal Information
Benzonase® Nuklease wird von der Merck KGaA, Darmstadt, Deutschland und/oder ihrer Tochtergesellschaften geliefert.
Benzonase is a registered trademark of Merck KGaA, Darmstadt, Germany
Lagerklasse
10 - Combustible liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves
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P Friedhoff et al.
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Overproduction of the extracellular Serratia marcescens nuclease in Escherichia coli results in aggregation and sequestration of a large amount of the protein in inclusion bodies. Only a relatively small amount is secreted into the medium from which it can be
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The field of proteomics is continually looking for new ways to investigate protein dynamics within complex biological samples. Recently, many researchers have begun to use RNA interference (RNAi) as a method of manipulating protein levels within their samples, but the ability to accurately determine these protein amounts remains a challenge. Fortunately, over the past decade, the field of proteomics has witnessed significant advances in the area of mass spectrometry. These advances, both in instrumentation and methodology, are providing researchers with sensitive assays for both identification and quantification of proteins within complex samples. This discussion will highlight some of these methodologies, namely the use of Multiple Reaction Monitoring (MRM) and Protein-AQUA.
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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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