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/e5014
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
Sign In to View Organizational & Contract Pricing
Select a Size
5 KU
R 1,870.82
25 KU
R 6,418.02
About This Item
CAS Number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.54
Skip To
biological source
Serratia marcescens
Quality Level
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
Looking for similar products? Visit Product Comparison Guide
1 of 4
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.
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
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
Choose from one of the most recent versions:
Already Own This Product?
Find documentation for the products that you have recently purchased in the Document Library.
Jos J M Drabbels et al.
Blood, 118(19), e149-e155 (2011-09-21)
Microchimerism is defined by the presence of low levels of nonhost cells in a person. We developed a reliable method for separating viable microchimeric cells from the host environment. For flow cytometric cell sorting, HLA antigens were targeted with human
The involvement of tau in nucleolar transcription and the stress response
Maina M.B., et al.
Acta Neuropathologica Communications, 6(70) (2018)
Characterization of Laminins in Healthy Human Aortic Valves and a Modified Decellularized Rat Scaffold
Granath C, et al.
BioResearch Open Access, 9(1) (2020)
G N Eaves et al.
Journal of bacteriology, 85(2), 273-278 (1963-02-01)
Eaves, George N. (Wayne State University College of Medicine, Detroit, Mich.) and Charles D. Jeffries. Isolation and properties of an exocellular nuclease of Serratia marcescens. J. Bacteriol. 85:273-278. 1963.-The exocellular nuclease of Serratia marcescens, isolated by anion-exchange chromatography on diethylaminoethyl-Sephadex
Richik Nilay Mukherjee et al.
Molecular biology of the cell, 30(18), 2349-2357 (2019-07-19)
Endoplasmic reticulum (ER) tubules and sheets conventionally correspond to smooth and rough ER, respectively. The ratio of ER tubules-to-sheets varies in different cell types and changes in response to cellular conditions, potentially impacting the functional output of the ER. To
Articles
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.
This page lists nine frequently asked questions and answers about Benzonase® Nuclease.
Benzonase®endonuclease efficiently removes nucleic acid contaminants from viral production, crucial for cell and gene therapies and vaccines.
Benzonase® Nuclease for reducing host DNA in microbiome workflows and enhancing taxa identification.
Related Content
Find protein research tools to prepare, isolate, and analyze proteins. Organized by how to extract, protect, purify, enrich, modify, and quantify proteins.
Read an automated protocol for protein purification using PureProteome™ nickel magnetic beads on the AAW™ automated assay workstation and see results comparing manual vs automated runs.
The use of Benzonase® endonuclease can significantly reduce the levels of DNA by more than 100,000-fold while also reducing viscosity and protecting downstream equipment from DNA fouling. However, optimization strategies and DoE are critical when it comes to reducing DNA in your process. Setting up a DoE for your Benzonase® endonuclease application can help you find the optimal operation conditions that deliver the required DNA clearance from your process.
-
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-
Helpful?
-
-
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?
-
-
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?
-
-
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?
-
-
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?
-
-
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?
-
-
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?
-
Active Filters
Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.
Contact Technical Service