标签归档：rnase

RNase B（对照底物） |NEB酶试剂 New England Biolabs

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产品信息

RNase B（对照底物） |
 
RNase B（对照底物）是一种高甘露糖型糖蛋白（1-3），可用作切割 N-连接糖链的糖苷内切酶的阳性对照。RNase B（对照底物）具有单个 N-连接糖基化位点，非常适合用于 SDS-PAGE 凝胶电泳迁移实验测定。

产品来源

牛胰腺

产品类别:
Glycoprotein Substrates Products,
Proteome Analysis Products

应用:
Protein Digestion,
Protein Labeling,
Recombinant Glycoprotein Expression,

Glycoprotein Analysis

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • P7817S     -20    
        RNase B P7817SVIAL -20 1 x 0.05 ml 5 mg/ml

  • 特性和用法

    贮存溶液

    50 µl Milli-Q water
    pH 7 @ 25°C

    分子量

    理论上的: 17000 道尔顿

  • 相关产品

    相关产品

    • p0702-endo-h
    • p0703-endo-hf
    • p0704-pngase-f
    • p0705-pngase-f-glycerol-free

  • 参考文献

    1. Plummer, T.H. and Hirs, C. (1964). J. Biol. Chem. 239, 2530-2538.
    2. Plummer, T.H. Jr., Tarentino, A. and Maley, F. (1968). J. Biochem. (Tokyo). 243, 5158-5164.
    3. Liang, C.-J. Yamashita, K. and Kobata, A. (1980). J. Biochem (Tokyo). 88, 51-58.

操作说明、说明书 & 用法

  • 操作说明

    1. RNase B Deglycosylation Protocol (P7817)

FAQs & 问题解决指南

  • FAQs

    1. Do detergents inhibit exoglycosidases/endoglycosidases?

  • 实验技巧

    这是糖苷内切酶 H、糖苷内切酶 Hf、PNGase F 和 PNGase F（无甘油）的很适合的阳性对照
    250 µg 足以进行大约 20 次反应

小鼠 RNase 抑制剂 |NEB酶试剂 New England Biolabs

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上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

小鼠 RNase 抑制剂是鼠源重组蛋白，分子量为 50 kDa。该抑制剂可以特异性抑制 RNase A、B 和 C 活性，它与 RNase 以 1:1 的比例高效非共价结合从而抑制其活性。但对 RNase 1、RNase T1、S1 核酸酶、RNase H 或来源于曲霉属（Aspergillus）的 RNase 无效。此外，与下列聚合酶共同使用时，该抑制剂不会抑制聚合酶的活性，如：Taq DNA 聚合酶、AMV 或 M-MuLV 反转录酶或噬菌体 RNA 聚合酶（SP6、T7 或 T3）。

重组小鼠 RNase 抑制剂不含半胱氨酸。已证实，人源 RNase 抑制剂中的半胱氨酸能导致抑制剂对氧化敏感甚至失活（1）。因此，小鼠 RNase 抑制剂与人/猪 RNase 抑制剂相比，抗氧化能力显著提高，且在 DTT 浓度较低（＜1 mM）时也很稳定。这使其在不适宜高浓度 DTT 的反应中更具优势（如： RT-PCR）。

产品来源

携带有鼠源 RNase 抑制剂基因的大肠杆菌菌株。

产品类别:
RNase Inhibitors,
PURExpress,
Cell-Free Protein Expression Products,

cDNA Synthesis & Reverse Transcriptases Products,
RNA Synthesis In vitro Transcription (IVT),

Protein Expression Products

应用:
NEBExpress® Cell-free E. coli Protein Synthesis System,
PURExpress,
Cell-Free Protein Expression,

Protein Expression,

PCR

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • M0314S     -20    
        RNase Inhibitor, Murine M0314SVIAL -20 1 x 0.075 ml 40,000 units/ml
    • M0314L     -20    
        RNase Inhibitor, Murine M0314LVIAL -20 1 x 0.375 ml 40,000 units/ml

  • 特性和用法

    单位定义

    1 单位指能抑制 5 ng RNase A 50%活性所需的小鼠 RNase 抑制剂的量。活性测定是通过抑制 RNase A 对胞嘧啶 2´，3´ -环单磷酸盐的水解来进行。

    贮存溶液

    20 mM HEPES-KOH
    50 mM KCl
    8 mM DTT
    50% Glycerol
    pH 7.6 @ 25°C

  • 优势和特性

    应用特性

    • RT-PCR
    • cDNA 合成
    • 体外转录/翻译
    • 酶催化的 RNA 标记反应
    • 需要完整 RNA 存在的其它应用

  • 注意事项

    1. 由于核糖核酸酶在变性条件下仍能保持活性，因此必须注意避免已与核糖核酸酶形成复合物的 RNase 抑制剂分子变性。为防止活性核糖核酸酶的释放，应避免反应温度超过 50℃和应用高浓度的尿素或其它变性剂。
    2. 反应中建议 RNase 抑制剂的浓度为 1 unit/μl。在建立反应时，RNase 抑制剂应先于其它可能作为 RNase 污染源的组份（如：酶、小提的质粒）加入到反应中。

  • 参考文献

    1. Kim, B.M. et al. Protein Science. 8, 430-434.

操作说明、说明书 & 用法

  • 操作说明

    1. Protocol for Avoiding Rnase Contamination using Murine Rnase Inhibitor (M0314)
    2. Poly(A) Tailing of RNA using E. coli Poly(A) Polymerase (NEB# M0276)

  • 应用实例

    • Scaling of High-Yield In vitro Transcription Reactions for Linear Increase of RNA Production

FAQs & 问题解决指南

  • 实验技巧

    该抑制剂的分子量为 50 kDa。它会在使蛋白质变性的条件下失活。

    我们建议在您的反应中抑制剂的用量为 1U/ul。

    抑制剂应先于其它可能成为 RNase 污染源的组份前加入反应。

    抑制剂应在低于 50℃的条件下使用。

DNase I (RNase-free) |NEB酶试剂 New England Biolabs

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上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

DNase I, (RNase-free) is an endonuclease that nonspecifically cleaves DNA to release di-, tri- and oligonucleotide products with 5´-phosphorylated and 3´-hydroxylated ends (1,2). DNase I acts on single- and double-stranded DNA, chromatin and RNA:DNA hybrids.

重点

Isolated from a recombinant source
Supplied with 10X Reaction Buffer

产品来源

An E. Coli strain that carries an MBP fusion clone of Bovine Pancreatic DNase I.

产品类别:
Exonucleases and Non-specific Endonucleases Products,
RNA Synthesis In vitro Transcription (IVT)

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • M0303S     -20    
        DNase I (RNase-free) M0303SVIAL -20 1 x 0.5 ml 2,000 units/ml
        DNase I Reaction Buffer B0303SVIAL -20 1 x 1.5 ml 10 X
    • M0303L     -20    
        DNase I (RNase-free) M0303LVIAL -20 2 x 1.25 ml 2,000 units/ml
        DNase I Reaction Buffer B0303SVIAL -20 1 x 1.5 ml 10 X

  • 特性和用法

    单位定义

    One unit is defined as the amount of enzyme which will completely degrade 1 µg of pBR322 DNA in 10 minutes at 37°C in DNase I Reaction Buffer.

    Complete degradation is defined as the reduction of the majority of DNA fragments to tetranucleotides or smaller.

    反应条件

    1X DNase I Reaction Buffer
    Incubate at 37°C

    1X DNase I Reaction Buffer
    10 mM Tris-HCl
    2.5 mM MgCl2
    0.5 mM CaCl2
    (pH 7.6 @ 25°C)

    使用浓度

    2,000 units/ml

    贮存溶液

    10 mM Tris-HCl
    2 mM CaCl2
    50% Glycerol
    pH 7.6 @ 25°C

    热失活

    75°C for 10 minutes

  • 优势和特性

    应用特性

    • Degradation of DNA template in transcription reactions
    • Removal of contaminating genomic DNA from RNA samples
    • DNase I footprinting
    • Nick Translation

  • 相关产品

    单独销售的组分

    • DNase I 反应缓冲液

  • 注意事项

    1. EDTA should be added to a final concentration of 5 mM to protect RNA from being degraded during enzyme inactivation (3).
    2. We do not recommend using NEB's DNase I as a substitute for Monarch DNase I in the RNA isolation workflow when using the Monarch Total RNA Miniprep Kit (NEB #T2010).  Monarch DNase I is optimized for use in this workflow, and is available for purchase as a component of the Monarch Total RNA Enzyme Pack, (NEB #T2019).

  • 参考文献

    1. Kunitz, M. (1950). J. Gen. Physiol . 33, 349-362.
    2. Vanecko, S. and laskowski, M. (1961). J. Biol. Chem . 236, 3312-3316.
    3. Huang, Z. et al. (1996). Biotechniques . 20, 1012-1020.

操作说明、说明书 & 用法

  • 操作说明

    1. A Typical DNase I Reaction Protocol (M0303)

  • 使用指南

    • Activity of DNA Modifying Enzymes in rCutSmart™ Buffer

工具 & 资源

  • 选择指南

    • Activities of Exonucleases and Non-specific Endonucleases
    • Common Applications for Exonucleases and Endonucleases
    • Properties of Exonucleases and Non-specific Endonucleases

  • Web 工具

    • Exo Selector

FAQs & 问题解决指南

  • FAQs

    1. What is the specific activity of DNase I(RNase-free)?
    2. Will DNase I work in NEB buffers 1-4?
    3. What is the best way to remove DNase I from my reaction?
    4. Will DNase I work in rCutSmart buffer?
    5. Can I use the Monarch RNA Cleanup Kit to cleanup up my DNase I-treated RNA?
    6. Can I use NEB DNase I (#M0303) with the Monarch Total RNA Miniprep Kit?

RNase Inhibitor, Human Placenta |NEB酶试剂 New England Biolabs

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上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

RNase Inhibitor, Human Placenta is a recombinant human placental protein which specifically inhibits ribonucleases (RNases) A, B and C (1). It is not effective against RNase 1, RNase T1, S1 Nuclease, RNase H or RNase from Aspergillus. In addition, no inhibition of polymerase activity is observed when RNase Inhibitor, Human Placenta is used with Taq DNA Polymerase, AMV or M-MuLV Reverse Transcriptases, or Phage RNA Polymerases (SP6, T7, or T3).

The 50 kDa protein inhibits RNases by binding noncovalently in a 1:1 ratio with an association constant greater than 1014 (2).

产品来源

An E. coli strain that carries the Ribonuclease Inhibitor gene from human placenta.

产品类别:
RNase Inhibitors,
RNA Synthesis In vitro Transcription (IVT)

应用:
RT-PCR & cDNA Synthesis

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • M0307S     -20    
        RNase Inhibitor, Human Placenta M0307SVIAL -20 1 x 0.05 ml 40,000 units/ml
    • M0307L     -20    
        RNase Inhibitor, Human Placenta M0307LVIAL -20 1 x 0.25 ml 40,000 units/ml

  • 特性和用法

    单位定义

    One unit is defined as the amount of RNase Inhibitor, Human Placenta required to inhibit the activity of 5 ng of RNase A by 50%. Activity is measured by the inhibition of hydrolysis of cytidine 2, 3′-cyclic monophosphate by RNase A.

    贮存溶液

    50 mM KCl
    8 mM DTT
    50% Glycerol
    20 mM HEPES-KOH
    pH 7.6 @ 25°C

  • 注意事项

    1. Since ribonucleases typically retain activity under denaturing conditions, care must be taken to avoid denaturing RNase Inhibitor molecules which have complexed with a ribonuclease. To prevent the release of active ribonuclease, temperatures greater than 50°C and high concentrations of urea or other denaturing agents should be avoided.
    2. The recommended concentration of RNase Inhibitor in a reaction is 1 unit/μl. During assembly of a reaction, RNase Inhibitor should be added before other components that are possible sources of RNase contamination (i.e. enzymes, plasmid from a mini prep.)

  • 参考文献

    1. Blackburn, P. and Moore, S. (1982). Pancreatic Ribonucleases . The Enzymes. XV, Part B, Academic Press, NY.
    2. Blackburn, P., Wilson, G. and Moore, S. (1977). J. Biol. Chem. 252,5904.

操作说明、说明书 & 用法

  • 操作说明

    1. Protocol for Avoiding Rnase Contamination using Rnase Inhibitor, Human Placenta (M0307)