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
Sinônimo(s):
Endonuclease
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Selecione um tamanho
5 KU
R$ 540,00
25 KU
R$ 1.851,00
Sobre este item
Número CAS:
UNSPSC Code:
12352204
NACRES:
NA.54
MDL number:
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Estamos aqui para ajudarNome do produto
Benzonase® Nuclease, ≥250 units/μL, ≥90% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution
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
Quality Level
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Este 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 |
Legal Information
A nuclease Benzonase® é fornecida pela Merck KGaA, Darmstadt, Alemanha e/ou suas filiais.
Benzonase is a registered trademark of Merck KGaA, Darmstadt, Germany
Other Notes
Uma unidade digere DNA de esperma de salmão sonicado em oligonucleotídeos solúveis em ácido equivalentes a ΔA260 de 1,0 em 30 min. Em pH 8,0 a 37 °C (volume de reação 2,625 ml).
Physical form
Solução em glicerol 50% contendo 20 mM de Tris HCl, pH 8,0, 2 mM de MgCl2 e 20 mM de NaCl.
Application
A nuclease Benzonase® tem sido utilizada: como componente no tampão de lise gelado C para digerir DNA e RNA, facilitando a liberação completa de todas as proteínas nucleares[1] na etapa de imunoprecipitação.Para liberar complexos proteicos do nucleoplasma e cromatina[2]como suplemento no RIPA para fracionar células SHSY5Y para imunoprecipitação[3] para remover ácidos nucleicos residuais das raízes aórticas no método de descelularização[4]
Usado para remoção de ácidos nucleicos de amostras proteicas.
Biochem/physiol Actions
A nuclease Benzonase® pode digerir completamente os ácidos nucleicos em oligonucleotídeos terminados em 5′-monofosfato com 3 a 5 bases de comprimento, tornando-se a ferramenta ideal para remover ácidos nucleicos de proteínas recombinantes e para aplicações que exigem a digestão completa de ácidos nucleicos. Além de reduzir a viscosidade dos extratos de proteína e evitar a aglutinação de células, o pré-tratamento de amostras de proteína com a nuclease Benzonase® pode melhorar significativamente sua resolução na eletroforese em gel 2D, eliminando todos os ácidos nucleicos ligados. Essa enzima versátil pode digerir DNA e RNA nativos ou desnaturados pelo calor, e seu pH ideal para a atividade enzimática é de 8,0 a 9,2. A nuclease benzonase® é eficaz na remoção do DNA do hospedeiro de amostras de microbioma. Em muitos casos, as amostras de microbioma (como saliva ou pele) terão uma alta porcentagem de DNA do hospedeiro que interfere nos resultados posteriores. Nossos especialistas mostram que a redução do DNA hospedeiro diminui o custo do sequenciamento, ao mesmo tempo em que aumenta e melhora os dados. Os dados experimentais são mostrados no artigo técnico - Nuclease Benzonase® para fluxos de trabalho de microbioma
Digere DNA e RNA nativos ou desnaturados por calor.
Features and Benefits
- Depleção de DNA hospedeiro em amostras de microbioma.
- Digestão eficaz de ácidos nucleicos em uma variedade de fluxos de trabalho.
- Redução da viscosidade durante a extração de proteínas.
General description
A nuclease Benzonase® é uma endonuclease altamente eficiente e geneticamente modificada, originária da Serratia marcescens[5]. Essa proteína dimérica, com duas ligações dissulfeto essenciais, [6] é capaz de atacar e degradar todas as formas de DNA e RNA (fita simples, fita dupla, linear e circular) em uma ampla gama de condições operacionais. A nuclease Benzonase® é capaz de remover ácidos nucleicos e melhorar a pureza e qualidade das amostras de proteína.
Classe de armazenamento
10 - Combustible liquids
wgk
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves
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Sequential fractionation and isolation of subcellular proteins from tissue or cultured cells
Baghirova S, et al.
MethodsX, 2, 440-445 (2015)
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)
P Friedhoff et al.
Protein expression and purification, 5(1), 37-43 (1994-02-01)
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
Miles C Scotcher et al.
PloS one, 4(3), e4924-e4924 (2009-03-18)
Botulism, an often fatal neuroparalytic disease, is caused by botulinum neurotoxins (BoNT) which consist of a family of seven serotypes (A-H) produced by the anaerobic bacterium Clostridium botulinum. BoNT, considered the most potent biological toxin known, is a 150 kDa
Artigos
Benzonase® Nuclease for reducing host DNA in microbiome workflows and enhancing taxa identification.
This discussion will highlight some of these methodologies, namely, the use of Multiple Reaction Monitoring (MRM) and Protein-AQUA.
Benzonase®endonuclease efficiently removes nucleic acid contaminants from viral production, crucial for cell and gene therapies and vaccines.
This page lists nine frequently asked questions and answers about Benzonase® Nuclease.
Conteúdo relacionado
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.
Find protein research tools to prepare, isolate, and analyze proteins. Organized by how to extract, protect, purify, enrich, modify, and quantify proteins.
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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-
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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?
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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