Pierce™ Universal Nuclease for Cell Lysis
Pierce™ Universal Nuclease for Cell Lysis
実際の製品は異なる場合があります
Thermo Scientific™

Pierce™ Universal Nuclease for Cell Lysis

Thermo Scientific Pierce Universal Nuclease for Cell Lysisは、細胞ライセートの調製時に核酸を完全に分解する必要があるさまざまなアプリケーションに最適です。Universal詳細を見る
製品番号(カタログ番号)数量
8870125 kU
887005 kU
88702100 kU
製品番号(カタログ番号) 88701
価格(JPY)
55,800
Each
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数量:
25 kU
一括またはカスタム形式をリクエストする
Thermo Scientific Pierce Universal Nuclease for Cell Lysisは、細胞ライセートの調製時に核酸を完全に分解する必要があるさまざまなアプリケーションに最適です。

Universal Nuclease for Cell Lysisの特長:

広範囲—あらゆる形態のDNAおよびRNAを分解
非常に高品質の酵素—SDS-PAGEによる試験で、ヌクレアーゼの純度は≥99%
高い活性—DNase Iより100倍高い比活性
多用途—幅広い細胞溶解試薬に使用可能

Pierce Universal Nuclease for Cell Lysisは、Serratia marcescensに由来する遺伝子組換えエンドヌクレアーゼです。この酵素は大腸菌から産生および精製され、2つの重要なジスルフィド結合を持つ2つの同一の30 kDaサブユニットで構成されています。このユニバーサルエンドヌクレアーゼは、1本鎖、2本鎖、線状、および環状DNAおよびRNAを分解し、幅広い温度とpHで効果を発揮できます。この酵素は比活性が高く(DNase Iの100倍)、他のヌクレアーゼよりも熱安定性が向上しています。Pierce Universal Nucleaseは純度≥99%の酵素で、測定可能なプロテアーゼ活性はなく、250 U/µLの濃度で提供されます。Pierce Universal Nuclease for Cell Lysisの性能は、Benzonase™ Nuclease(EMD Merck)と同等です。

アプリケーション:
• B-PER、Y-PER、またはその他の市販および自家製細胞溶解試薬や機械的破壊と併用して、タンパク質抽出物の粘度を低減
• ダウンストリーム処理の前に組換えタンパク質調製でDNAとRNAを除去

Pierce Universal Nuclease for Cell Lysisは一般的に、タンパク質調製物から核酸を除去することにより、ダウンストリームアプリケーション用に細菌や哺乳類のタンパク質抽出物の粘度を下げるために使用します。この酵素は、塩基長が5未満のオリゴヌクレオチドにまで核酸を完全に分解します。Pierce Universal Nuclease for Cell Lysisは、ライセートペレットの上清からの分離を向上させ、処理済みライセートのろ過を増強します。また、クロマトグラフィーの処理時間を短縮し、タンパク質の全体的な収量を増大させます。このエンドヌクレアーゼは、タンパク質抽出物の2Dゲル電気泳動への適合性を向上させることも実証されています。1ユニットは、ニシンDNAを37℃で30分間超音波処理して、260 nmでの吸光度を1.0変化させるのに必要な酵素量に匹敵します(標準ヌクレアーゼを使用して、Merck™ Serratia marcescens体積活性アッセイにより決定)。

関連製品
Micrococcal Nuclease Solution (≥ 1 unit/µL)
研究用途にのみご使用ください。診断目的には使用できません。
仕様
バッファーLysis Buffer
酵素ヌクレアーゼ
数量25 kU
試薬タイプ細胞溶解用酵素
形状液体
製品ラインPierce
製品タイプCell Lysis Enzyme
Unit SizeEach
組成および保存条件
直射日光を避け、涼しく乾燥した換気の良い場所に保存。

よくあるご質問(FAQ)

After cell lysis, my mass spectrometry sample is very viscous and difficult to pipette. How do I reduce the sample viscosity?

High sample viscosity after lysis is due to release of DNA from the nucleus. Sonication or addition of a nuclease such as the Pierce Universal Nuclease (Cat. No. 88700, 88701, or 88702) can be used to degrade DNA and reduce sample viscosity.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.

Can you explain how the B-PER Bacterial Protein Extraction Reagent lyses cells?

The B-PER Reagent solution contains a proprietary, mild, non-ionic detergent in 20 mM Tris-HCl, pH 7.5. It effectively disrupts cells and solubilizes native or recombinant proteins without denaturation. The reagent creates holes in the cell membrane that will leak out cytosolic proteins. The sample may become very viscous when the bacterial chromosome is released. We recommend adding DNAse I (Cat. No. 90083) to the reagent to reduce viscosity. For better lysis efficiency and if there are inclusion bodies, we recommend adding Lysozyme (Cat. No. 90082) to the reagent. Alternatively, you may purchase the B-PER Bacterial Protein Extraction Reagent with Enzymes Kit (Cat. No. 90078 or 90079) that includes the B-PER Bacterial Protein Extraction Reagent, DNase I, and Lysozyme.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

引用および参考文献 (5)

引用および参考文献
Abstract
Validation of an IFN-gamma ELISpot assay to measure cellular immune responses against viral antigens in non-human primates.
Authors:Yang F,Patton K,Kasprzyk T,Long B,Gupta S,Zoog SJ,Tracy K,Vettermann C
Journal:Gene therapy
PubMed ID:33432123
Adeno-Associated Virus (AAV)-based gene therapy vectors are in development for many inherited human disorders. In nonclinical studies, cellular immune responses mediated by cytotoxic T cells may target vector-transduced cells, which could impact safety and efficacy. Here, we describe the bioanalytical validation of an interferon-gamma (IFN-γ)-based Enzyme-Linked Immunospot (ELISpot) assay for ... More
Characterization of an expanded set of assays for immunomodulatory proteins using targeted mass spectrometry.
Authors:Whiteaker JR,Zhao L,Schoenherr RM,Huang D,Kennedy JJ,Ivey RG,Lin C,Lorentzen TD,Colantonio S,Caceres TW,Roberts RR,Knotts JG,Reading JJ,Perry CD,Garcia-Buntley SS,Bocik W,Hewitt SM,Paulovich AG
Journal:Scientific data
PubMed ID:38918394
Immunotherapies are revolutionizing cancer care, but many patients do not achieve durable responses and immune-related adverse events are difficult to predict. Quantifying the hundreds of proteins involved in cancer immunity has the potential to provide biomarkers to monitor and predict tumor response. We previously developed robust, multiplexed quantitative assays for ... More
PhoXplex: Combining Phospho-enrichable Cross-Linking with Isobaric Labeling for Quantitative Proteome-Wide Mapping of Protein Interfaces.
Authors:Hoenger Ramazanova RD,Roumeliotis TI,Wright JC,Choudhary JS
Journal:Journal of proteome research
PubMed ID:39422127
Integrating cross-linking mass spectrometry (XL-MS) into structural biology workflows provides valuable information about the spatial arrangement of amino acid stretches, which can guide elucidation of protein assembly architecture. Additionally, the combination of XL-MS with peptide quantitation techniques is a powerful approach to delineate protein interface dynamics across diverse conditions. While ... More
An efficient and cost-effective purification protocol for Staphylococcus aureus Cas9 nuclease.
Authors:Teng ACT,Tavassoli M,Shrestha S,Marks RM,McFadden MJ,Evagelou SL,Lindsay K,Vandenbelt A,Li W,Ivakine E,Cohn R,Santerre JP,Gramolini AO
Journal:STAR protocols
PubMed ID:36574341
Here, we describe a protocol for purifying functional clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) from Staphylococcus aureus within 24 h and over 90% purity. SaCas9 purification begins with immobilized metal affinity chromatography, followed by cation exchange chromatography, and ended with centrifugal concentrators. The simplicity, cost-effectiveness, and reproducibility ... More
p53 is active in murine stem cells and alters the transcriptome in a manner that is reminiscent of mutant p53.
Authors:Yan H, Solozobova V, Zhang P, Armant O, Kuehl B, Brenner-Weiss G, Blattner C
Journal:
PubMed ID:25719246
Since it was found that p53 is highly expressed in murine embryonic stem cells, it remained a mystery whether p53 is active in this cell type. We show that a significant part of p53 is localised in the nucleus of murine embryonic stem cells and that the majority of this ... More