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Merck

E1014

Benzonase®核酸酶

≥250 units/μL, ≥90% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution

Benzonase® Nuclease

同義詞:

内切核酸酶 来源于粘质沙雷氏菌

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CAS 編號:
MDL號碼:
分類程式碼代碼:
12352204
NACRES:
NA.54

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生物源

Serratia marcescens

品質等級

重組細胞

expressed in E. coli

化驗

≥90% (SDS-PAGE)

形狀

buffered aqueous glycerol solution

分子量

30 kDa

濃度

≥250 units/μL

應用

research use

異物活動

protease, essentially free

運輸包裝

wet ice

儲存溫度

−20°C

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本產品
706647120671205-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

一般說明

Benzonase®全能核酸酶是一种通过高效基因工程法从粘质沙雷氏菌(Serratia marcescens)中获得的核酸内切酶[1]。这种由两个必需二硫键连接的二聚体蛋白[2]可在宽泛的操作条件下攻击并降解所有形式的DNA和RNA(单链、双链、线性和环化)。Benzonase®全能核酸酶能够去除核酸并提高蛋白质样本的纯度和质量。

應用

Benzonase®全能核酸酶用于:作为冰冷裂解缓冲液C的组分以消化DNA 和  RNA,便于充分释放所有核蛋白[3]在免疫沉淀中用于释放核质和染色质中的蛋白复合物[4]作为RIPA添加剂,用于分离SHSY5Y细胞进行免疫沉淀[5]在脱细胞方法中用于去除主动脉根部的残留核酸[6]
用于去除蛋白样品中的核酸。

生化/生理作用

Benzonase®全能核酸酶能将核酸完全消化成长度为3到5个碱基的5′-单磷酸末端寡核苷酸,使其成为从重组蛋白中去除核酸的理想工具,并适于需要完全消化核酸的应用。除了降低蛋白质提取物的粘度并防止细胞结团之外,使用Benzonase®全能核酸酶预处理蛋白质样本可以消除任何结合的核酸,进而显著提高2D凝胶电泳的分辨率。这种多功能酶可消化天然或热变性的DNA和RNA,其酶活性的最佳pH值为8.0-9.2。Benzonase®全能核酸酶可有效去除微生物组样本中的宿主DNA。在许多情况下,微生物组样本(如唾液或皮肤等)中含有较高比例的能够干扰下游结果的宿主DNA。我们的专家表示,减少宿主DNA可以降低测序成本,同时增加并改进数据。实验数据显示于以下技术文章中-用于微生物组工作流程的Benzonase®全能核酸酶物
消化天然或热变性的DNA和RNA。

特點和優勢

  • 去除微生物组样本的宿主DNA。


  • 在各种工作流程中高效去除核酸。


  • 在蛋白提取过程中降低粘度。

外觀

在含有20 mM Tris HCl, pH 8.0, 2 mM MgCl2及20 mM NaCl的50%甘油中的溶液。

其他說明

在pH 8.0,37 °C条件下,一个单位的酶在30分钟内可将超声的鲑鱼精子DNA消化成等同于1.0 ΔA260的酸可溶寡核苷酸(2.625 ml反应体积)。

法律資訊

Benzonase®核酸酶由德国达姆施塔特默克公司和/或其附属公司提供。
Benzonase is a registered trademark of Merck KGaA, Darmstadt, Germany

儲存類別代碼

10 - Combustible liquids

水污染物質分類(WGK)

WGK 1

閃點(°F)

Not applicable

閃點(°C)

Not applicable

個人防護裝備

Eyeshields, Gloves


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分析證明 (COA)

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您可以在文件庫中找到最近購買的產品相關文件。

存取文件庫

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)
Sequential fractionation and isolation of subcellular proteins from tissue or cultured cells
Baghirova S, et al.
MethodsX, 2, 440-445 (2015)
DONSON facilitates Cdc45 and GINS chromatin association and is essential for DNA replication initiation
Kingsley G, et al.
Nucleic Acids Research, 51(18), 9748?9763-9748?9763 (2023)
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

文章

Benzonase®endonuclease efficiently removes nucleic acid contaminants from viral production, crucial for cell and gene therapies and vaccines.

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.

Benzonase® Nuclease for reducing host DNA in microbiome workflows and enhancing taxa identification.

This page lists nine frequently asked questions and answers about Benzonase® Nuclease.

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相關內容

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.

Questions

1–7 of 7 Questions  
  1. 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
    1. 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

      Helpful?

  2. 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
    1. 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?

  3. Hi, what is the ratio of E1014 : DNA and E1014 : RNA for an efficient acid nucleic removal?

    1 answer
    1. 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?

  4. How many ul does it contain? What kind of Unit is KU?

    1 answer
    1. 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-documentation

      Helpful?

  5. 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
    1. The recommended starting concentration for Benzonase nuclease is 25 units per milliliter of cell lysate.

      Helpful?

  6. 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
    1. 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?

  7. What is the storage temperature range for E1014? There is only a specified storage temperature, and not a range.

    1 answer
    1. This product is stored at freezer temperature, which is typically -20°C. An excepted range is -15 - -20°C.

      Helpful?

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