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.
E1014
Benzonase® ヌクレアーゼ
≥250 units/μL, ≥90% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution
Benzonase® Nuclease
別名:
エンドヌクレアーゼ Serratia marcescens（セラチア菌）由来
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この商品について
CAS番号:
UNSPSC Code:
12352204
NACRES:
NA.54
MDL number:
Assay:
≥90% (SDS-PAGE)
Biological source:
Serratia marcescens
Recombinant:
expressed in E. coli
Concentration:
≥250 units/μL
スキップする
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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当該品目 | 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 |
Application
Benzonase®ヌクレアーゼは、次のような用途で使用されています：DNA と RNAを消化してすべての核タンパク質の完全放出を促進する氷冷溶解バッファーCの成分として使用するため[1]、免疫沈降ステップにおいて、核質およびクロマチンからタンパク質複合体を放出するため[2]、RIPA法のサプリメントとして免疫沈降法のためにSHSY5Y細胞を分画するため[3]、脱細胞化法において大動脈根から残存する核酸を除去するため[4]
タンパク質サンプルから核酸を除去する際に使用します。
Biochem/physiol Actions
Benzonase®ヌクレアーゼは、核酸を5′-一リン酸終端を持つ長さ3～5塩基のオリゴヌクレオチドへと完全に消化できるため、組換えタンパク質からの核酸の除去や核酸の完全消化が必要なアプリケーションに最適なツールとなっています。タンパク質抽出物の粘度を低下させ、細胞凝集を予防することに加えて、Benzonase®ヌクレアーゼでタンパク質サンプルを事前処理することで、結合している核酸を除去することにより2Dゲル電気泳動における分離を著しく改善できます。この汎用性の高い酵素は、未変性と熱変性の両方のDNAとRNAを消化することができ、酵素活性至適pHは8.0～9.2であることが確認されています。Benzonase® ヌクレアーゼは、マイクロバイオームサンプルから宿主DNAを効果的に除去します。多くの場合、マイクロバイオームサンプル（唾液や皮膚など）には、宿主DNAが高い割合で含まれており、これがダウンストリームの結果を妨げます。メルクのエキスパートは、宿主DNAの低減により、データを増やし改善しながらシーケンシング費用を削減できることを明らかにしています。実験データは、技術論文『Benzonase® Nuclease for Microbiome Workflows』に掲載しています。
未変性または熱変性のDNA, RNAを分解します。
Features and Benefits
- マイクロバイオームサンプル中の宿主DNAの除去。
- さまざまなワークフローにおける効果的な核酸消化。
- タンパク質抽出時の粘度の低減。
General description
Legal Information
ベンゾナーゼ®ヌクレアーゼは、Merck KGaA（ダルムシュタット、ドイツ）および/またはその関連会社が供給しています。
Benzonase is a registered trademark of Merck KGaA, Darmstadt, Germany
Other Notes
1ユニットは、反応液を2.625 mLとした場合に、pH 8.0、37°C、30分間で、超音波破砕されたサケ精子DNAを切断して、1.0のΔA260に相当する酸可溶性オリゴヌクレオチドを生じる酵素量です。
Physical form
50%グリセロ-ル溶液(20 mM Tris-HCl(pH 8.0), 2 mM MgCl2, 20 mM NaClを含有)。
保管分類
10 - Combustible liquids
wgk
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves
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
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
資料
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® Nuclease for reducing host DNA in microbiome workflows and enhancing taxa identification.
このページは、Benzonase®（ベンゾナーゼ）ヌクレアーゼについてよくある質問9項目とその回答をまとめたものです。
関連コンテンツ
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.
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.
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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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 回答-
役に立ちましたか？
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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 回答-
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役に立ちましたか？
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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 回答-
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.
役に立ちましたか？
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Hi, what is the ratio of E1014 : DNA and E1014 : RNA for an efficient acid nucleic removal?
1 回答-
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.
役に立ちましたか？
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How many ul does it contain? What kind of Unit is KU?
1 回答-
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役に立ちましたか？
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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 回答-
The recommended starting concentration for Benzonase nuclease is 25 units per milliliter of cell lysate.
役に立ちましたか？
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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 回答-
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.
役に立ちましたか？
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What is the storage temperature range for E1014? There is only a specified storage temperature, and not a range.
1 回答-
This product is stored at freezer temperature, which is typically -20°C. An excepted range is -15 - -20°C.
役に立ちましたか？
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