Sodium dodecyl sulfate concentrations up to 0.05% do not affect enzyme activity. Concentrations of 0.1% or 1% reduce activity more substantially because sodium dodecyl sulfate denatures the enzyme.
At sodium deoxycholate concentrations below 0.4%, Benzonase® endonuclease retains at least 70% activity. At a concentration of 1%, activity is approximately 30%.
Magnesium or manganese ions at concentrations of 1–2 millimolar are essential for activity. Magnesium ions are preferred because they enable the enzyme to reach optimal activity.
E1014
Benzonase® Nuclease
≥250 units/μL, ≥90% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution
Synonym(s):
Endonuclease from Serratia marcescens
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About This Item
CAS Number:
UNSPSC Code:
12352204
NACRES:
NA.54
EC Number:
MDL number:
Assay:
≥90% (SDS-PAGE)
Biological source:
Serratia marcescens
Recombinant:
expressed in E. coli
Concentration:
≥250 units/μL
Pricing and availability is not currently available.
biological source
Serratia marcescens
Quality Segment
recombinant
expressed in E. coli
product line
Benzonase®
assay
≥90% (SDS-PAGE)
form
buffered aqueous glycerol solution
mol wt
30 kDa
concentration
≥250 units/μL
input
general use sample(s)
application(s)
research use
foreign activity
protease, essentially free
shipped in
wet ice
storage temp.
−20°C
General description
Benzonase® nuclease is a highly efficient and genetically engineered endonuclease that originates from Serratia marcescens. This dimeric protein with two essential disulfide bonds is capable of attacking and degrading all forms of DNA and RNA (single-stranded, double-stranded, linear, and circular) under a wide range of operating conditions. Benzonase® nuclease is capable of removing nucleic acids and enhancing the purity and quality of protein samples.
Application
Used for the removal of nucleic acid from protein samples.
Benzonase® Nuclease has been used: as a component in ice-cold lysis buffer C to digest DNA and RNA to facilitate the complete release of all nuclear proteins in the immunoprecipitation step to release protein complexes from the nucleoplasm and chromatinas a supplement in RIPA to fractionate SHSY5Y cells for immunoprecipitation to remove residual nucleic acids from the aortic roots in decellularization method
Biochem/physiol Actions
Benzonase® Nuclease can completely digest nucleic acids into 5′-monophosphate terminated oligonucleotides of 3 to 5 bases in length, making it the ideal tool for removing nucleic acids from recombinant proteins and for applications that require complete digestion of nucleic acids. In addition to reducing viscosity in protein extracts and preventing cell clumping, pretreatment of protein samples with Benzonase® nuclease can significantly improve their resolution on 2D gel electrophoresis by eliminating any bound nucleic acids. This versatile enzyme can digest both native or heat-denatured DNA and RNA, with its optimum pH for enzyme activity found to be 8.0-9.2. Benzonase® nuclease is effective at removing host DNA from microbiome samples. In many cases, microbiome samples (such as saliva or skin) will have a high percentage of host DNA that interferes with downstream results. Our experts show that the reduction of host DNA lowers the cost of sequencing while increasing and improving the data. Experimental data is shown in the technical article - Benzonase® Nuclease for Microbiome Workflows
Digests native or heat-denatured DNA and RNA.
Features and Benefits
- Host DNA depletion in microbiome samples.
- Effective nucleic acid digestion in a variety of workflows.
- Viscosity reduction during protein extraction.
Physical form
Solution in 50% glycerol containing 20 mM Tris HCl, pH 8.0, 2 mM MgCl2, and 20 mM NaCl.
Other Notes
One unit will digest sonicated salmon sperm DNA to acid-soluble oligonucleotides equivalent to a ΔA260 of 1.0 in 30 min at pH 8.0 at 37 °C (reaction volume 2.625 ml).
Legal Information
Benzonase® Nuclease is supplied by Merck KGaA, Darmstadt, Germany and/or its affiliates.
Benzonase is a registered trademark of Merck KGaA, Darmstadt, Germany
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This Item | |||
|---|---|---|---|
| description ≥250 units/μL, ≥90% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution | description - | description ≥250 units/μL, ≥99% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution, ultrapure grade | description - |
| biological source Serratia marcescens | biological source Serratia marcescens | biological source Serratia marcescens | biological source donkey |
| assay ≥90% (SDS-PAGE) | assay >90% (SDS-PAGE) | assay ≥99% (SDS-PAGE) | assay - |
| application(s) research use | application(s) research use | application(s) research use | application(s) research pathology |
| concentration ≥250 units/μL | concentration 25-29 units/μL | concentration ≥250 units/μL | concentration - |
| recombinant expressed in E. coli | recombinant expressed in E. coli | recombinant expressed in E. coli | recombinant - |
| form buffered aqueous glycerol solution | form buffered aqueous glycerol solution | form buffered aqueous glycerol solution | form - |
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Storage Class
10 - Combustible liquids
Flash Point (°F)
Not applicable
PPE (personal protective equipment)
Eyeshields, Gloves
Flash Point (°C)
Not applicable
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I am doing a protein extraction and would like to know if your nuclease would have activity in a RIPA buffer with 0.1% SDS and 1% sodium deoxycholate, without MgCl₂? Also, could you tell me the recommended incubation condition in this case?
1 answer-
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I want to use the Benzonase with a pH 7 buffer to remove DNA from the cell lysate. How significantly does the buffer pH affect the nuclease activity of Benzonase?
1 answer-
For assay purposes, this product is tested at pH 8.0 and has an optimum activity range of pH 8.0 - 9.0. However significant nuclease activity is generally retained at a pH range of 6.0 - 10.0.
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For a phoshpoproteomic sample prep for T cells (high DNA content), how long would you recommend the concentration and incubation time to be?
1 answer-
The unit activity of this product is defined as the amount of enzyme that causes an delta ΔA260 of 1.0 in 30 minutes, which corresponds to complete digestion of 37 ug of DNA. A general recommendation for 106 -107 cells is 1 -2 µl of Benzonase Nuclease along with 100 µl RIPA or the preferred lysis buffer and incubate for 30 minutes at room temperature. Increased incubation periods may be required for samples with high DNA content.
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Does this product have a tag (e.g His)
1 answer-
This item does not contain a His-tag.
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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-
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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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.
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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.
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How many ul does it contain? What kind of Unit is KU?
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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.
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