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.
E1014
Benzonase® Nuclease
≥250 units/μL, ≥90% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution
Benzonase® Nuclease
Synonym(s):
Endonuclease from Serratia marcescens
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5 KU
¥16,400
25 KU
¥53,600
About This Item
CAS Number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.54
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biological 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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This Item | 70664 | 71206 | 71205-M |
|---|---|---|---|
| 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® nuclease is a highly efficient and genetically engineered endonuclease that originates from Serratia marcescens[1]. This dimeric protein with two essential disulfide bonds [2] 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
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[3] in the immunoprecipitation step to release protein complexes from the nucleoplasm and chromatin[4]as a supplement in RIPA to fractionate SHSY5Y cells for immunoprecipitation[5] to remove residual nucleic acids from the aortic roots in decellularization method[6]
Used for the removal of nucleic acid from protein samples.
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.
Unit Definition
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).
Physical form
Solution in 50% glycerol containing 20 mM Tris HCl, pH 8.0, 2 mM MgCl2, and 20 mM NaCl.
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
Storage Class Code
10 - Combustible liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Personal Protective Equipment
dust mask type N95 (US), Eyeshields, Gloves
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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-
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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?
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.
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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.
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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.
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