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Phusion® HF 反应缓冲液套装 |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

NEB 公司提供专用的 5X 反应缓冲液和补充试剂，适用于 Thermo Scientific® Phusion 超保真 DNA 聚合酶及相关产品；缓冲液和试剂均由 NEB 公司生产并进行质量控制。

产品类别:
Phusion® High-Fidelity DNA Polymerases Products,
Reaction Buffers Products,
Buffers Products

应用:
Hot Start PCR,
Long Range PCR,
Fast PCR,

High-Fidelity PCR,
Specialty PCR,
Routine PCR,

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
    • B0518S     -20    
        Phusion® HF Buffer Pack B0518SVIAL -20 4 x 1.5 ml 5 X
        MgCl2 solution B0510AVIAL -20 2 x 1.5 ml 50 mM
        DMSO B0515AVIAL -20 1 x 0.5 ml 100 %

  • 相关产品

    相关产品

    • Phusion® GC 反应缓冲液套装
    • Phusion® High-Fidelity PCR Kit
    • m0530-phusion-high-fidelity-dna-polymerase
    • Phusion® 超保真 PCR 预混液（提供 HF 缓冲液）
    • Phusion® 超保真 PCR 预混液（提供 GC 缓冲液）
    • Phusion® Hot Start Flex DNA Polymerase
    • Phusion® 热启动 Flex 2X 预混液

操作说明、说明书 & 用法

  • 使用指南

    • Activity of Restriction Enzymes in PCR Buffers

工具 & 资源

  • Web 工具

    • Tm Calculator

FAQs & 问题解决指南

  • FAQs

    1. What are the differences between the various available Phusion® products?

Endo Hf |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

Endo Hf |
 

Endo Hf is a recombinant protein fusion of Endoglycosidase H and maltose binding protein. Endo Hf cleaves within the chitobiose core of high mannose and some hybrid oligosaccharides from N-linked glycoproteins (1) equally as well as Endo H.

Endo Hf |
Endo Hf |
60 μg of RNase B was incubated with 3,000 units of Endo H or Endo Hf under standard assay conditions
Endo Hf |
Aliquots were removed at various time points and measured for released carbohydrate. [Dubois et al. (1956) Anal. Chem. 28, 350-356].
Mobility Shift Analysis
Endo Hf |
1 unit of Endo H, Endo Hf or PNGase F was incubated per 10 µg of RNase B under standard assay conditions. At various time points, aliquots were removed and analyzed on a 10-20% SDS-gel for carbohydrate (CHO) release. 1 unit is defined as the amount of enzyme required to remove >95% of the carbohydrate from 10 µg of RNase B in one hour at 37°C.

产品来源

Cloned from Streptomyces picatus (2) and overexpressed in E.coli (3).

产品类别:
Endoglycosidases Products,
Proteome Analysis Products

应用:
Expression Systems,
Glycan Sequencing,
Proteomics,

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
    • P0703S     -20    
        Endo Hf P0703SVIAL -20 1 x 0.1 ml 1,000,000 units/ml
        GlycoBuffer 3 B1720SVIAL -20 1 x 1 ml 10 X
        Glycoprotein Denaturing Buffer B1704SVIAL -20 1 x 1 ml 10 X
    • P0703L     -20    
        Endo Hf P0703LVIAL -20 1 x 0.5 ml 1,000,000 units/ml
        GlycoBuffer 3 B1720SVIAL -20 1 x 1 ml 10 X
        Glycoprotein Denaturing Buffer B1704SVIAL -20 1 x 1 ml 10 X

  • 特性和用法

    单位定义

    One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 10 µg of denatured RNase B in 1 hour at 37°C in a total reaction volume of 10 µl (10 NEB units = 1 IUB milliunit). 

    Unit Definition Assay: 
    10 µg of RNase B are denatured with 1X Glycoprotein Denaturing Buffer (0.5% SDS, 40 mM DTT) at 100°C for 10 minutes. After the addition of 1X GlycoBuffer 3, two-fold dilutions of Endo Hf are added and the reaction mix is incubated for 1 hour at 37°C. Separation of reaction products are visualized by SDS-PAGE.

    1X Glycoprotein Denaturing Buffer
    0.5% SDS
    40 mM DTT

    反应条件

    1X GlycoBuffer 3
    Incubate at 37°C

    1X GlycoBuffer 3
    50 mM sodium acetate
    (pH 6 @ 25°C)

    贮存溶液

    20 mM Tris-HCl
    50 mM NaCl
    5 mM EDTA
    pH 7.5 @ 25°C

    热失活

    75°C for 10 minutes

    分子量

    实际: 70 kDa

  • 相关产品

    相关产品

    • p0702-endo-h
    • p0704-pngase-f
    • p0705-pngase-f-glycerol-free
    • RNase B（对照底物）
    • p0706-remove-it-pngase-f
    • Endo S
    • 糖苷内切酶 D

  • 注意事项

    1. Enzymatic activity is not affected by SDS.
    2. To deglycosylate a native glycoprotein, longer incubation time as well as more enzyme may be required.
    3. Activity at different temperatures (measured after a 1 hour incubation of glycosidase and denatured RNase B at the given temperature): 37°C – 100%; 30°C – 65%; 25°C – 40%; 17°C – 25% and 2°C – 0%.
    4. Typical reaction conditions: Please see the Protocol Tab.

  • 参考文献

    1. Maley, F. et al. (1989). Anal. Biochem. 180, 195-204.
    2. Robbins, P. et al. (1984). J. Biol. Chem. 259, 7577-7583.
    3. Guan, C. and Wong, S. New England Biolabs, unpublished Observations. Unpublished observation

操作说明、说明书 & 用法

  • 操作说明

    1. Endo H/Endo Hf Protocol
    2. Protocol for Endo H/Hf Non-Denaturing P0702 and P0703

工具 & 资源

  • 选择指南

    • Endoglycosidase Selection Chart

FAQs & 问题解决指南

  • FAQs

    1. What is the difference between PNGase F and Endo H?
    2. What is the difference between Endo H and Endo Hf?
    3. I tried the Endo H/Hf on my glycoprotein and it failed. What could be the problem?
    4. How much Endo H/Endo Hf should I use?
    5. Is EndoH/ Endo Hf inhibited by SDS?
    6. What are the typical reaction conditions for Endo Hf?
    7. Are Protease Inhibitors acceptable for use in an Endo H/Hf reaction?
    8. Why have the NEB Glycosidase enzymes changed reaction buffers? What are the new reaction buffers and can I still use an enzyme with its old buffer? Where can I find the composition of the old buffers?
    9. What is a good endoglycosidase substrate?
    10. Do detergents inhibit exoglycosidases/endoglycosidases?
    11. What are Glycosidases and their uses?

  • 实验技巧

    You can use this enzyme under native or denaturing conditions

    Under native conditions we recommend adding more enzyme and using longer incubation times

    Enzymatic activity is not affected by SDS

    A good positive control substrate is RNase B

915002-德国MN水质快速测试盒

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产品型号915002

品       牌德国MN

产品简介

德国MN水质快速测试盒 残留硬度测试盒（H2F）用于锅炉补给水、食品饮料行业、皮革化工、造纸工业、地表水和海水、纺织工业等，欢迎前来咨询！

详情介绍

残留硬度测试盒（H2F）/德国MN水质快速测试盒915002

残留硬度测试盒（H2F）/德国MN水质快速测试盒HE total Hardness H 2 F

残留硬度测试盒（H2F）

产品编号：915 002（915202）

测试类型： Titr(高灵敏度容量分析法测试盒)；

测试范围：1 syringe is sufficient for the determination of 0.05 – 2.0 °d or 0.01 – 0.36 mmol/l Ca2+(corresponds to 1 – 36 mg/l CaCO3).

测试次数：200次；

保质期：18个月；

应用 ：锅炉补给水、食品饮料行业、皮革化工、造纸工业、地表水和海水、纺织工业

SalI-HF® |NEB酶试剂 New England Biolabs

发表于由

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

高保真（HF）限制性内切酶在 rCutSmart 缓冲液中具有 100% 活性；统一缓冲液意味着更加直接、简化的样品处理过程。HF 内切酶还会显著降低星号活性。所有 HF 内切酶均符合省时酶（Time-Saver）标准，可在 5-15 分钟内酶切底物 DNA，也可实现过夜酶切而不会造成 DNA 降解。HF 限制性内切酶在改造时便将性能作为重要指标，可在更宽的条件范围下具有完全活性，最大限度地减少非特异性酶切产物，同时为实验设计提供灵活性。

NEB 所有常规和高保真（HF）限制性内切酶均经过严格的质量控制。部分 HF 限制性内切酶的 核酸酶污染研究如下所示。

限制性内切酶竞争品牌研究：核酸酶污染

SalI-HF® |


用多供应商的 EcoRI、NotI 和 BamHI，在含有荧光标记单链平末端、双链平末端、3’ 突出末端 或 5’ 突出末端的寡核苷酸中进行测试。 通过毛细管电泳和峰值分析确定降解百分比。分辨率达到 单核苷酸水平。

产品来源

大肠杆菌菌株，携带有克隆自白色链霉菌（Streptomyces albus）G（ATCC 49789）并经修饰的 SalI 基因

产品类别:
Methylation Sensitive Restriction Enzymes for Epigenetics Products,
Restriction Endonucleases S Products,
High-Fidelity (HF®) Restriction Endonucleases Products,

Time-Saver Qualified Restriction Enzymes Products

应用:
Restriction Enzyme Digestion

  • 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
    • R3138V     -20    
        SalI-HF® R3138VVIAL -20 1 x 0.05 ml 20,000 units/ml
        rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
        Gel Loading Dye, Purple (6X) B7024VVIAL 25 1 x 0.2 ml 6 X
    • R3138S     -20    
        SalI-HF® R3138SVIAL -20 1 x 0.1 ml 20,000 units/ml
        rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
        Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X
    • R3138T     -20    
        SalI-HF® R3138TVIAL -20 1 x 0.02 ml 100,000 units/ml
        rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
        Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X
    • R3138L     -20    
        SalI-HF® R3138LVIAL -20 1 x 0.5 ml 20,000 units/ml
        rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
        Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X
    • R3138M     -20    
        SalI-HF® R3138MVIAL -20 1 x 0.1 ml 100,000 units/ml
        rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
        Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X

  • 特性和用法

    单位定义

    一个单位是指在 50 μl 的总反应体系中，37℃ 下，1 小时内酶切 1 μg λ DNA（HindIII 酶切）所需的酶量。

    反应条件

    1X rCutSmart™ 缓冲液
    Incubate at 37°C

    1X rCutSmart™ 缓冲液
    50 mM Potassium Acetate
    20 mM Tris-acetate
    10 mM Magnesium Acetate
    100 µg/ml Recombinant Albumin
    (pH 7.9 @ 25°C)

    在不同缓冲液中的活性

    NEBuffer™ r1.1: 10%
    NEBuffer™ r2.1: 100%
    NEBuffer™ r3.1: 100%
    rCutSmart™ Buffer: 100%

    稀释兼容性

    • 稀释液 A

    贮存溶液

    10 mM Tris-HCl
    50 mM KCl
    1 mM DTT
    0.1 mM EDTA
    50% Glycerol
    300 µg/ml Recombinant Albumin
    pH 7.5 @ 25°C

    热失活

    65°C for 20 minutes

    甲基化敏感性

    dam 甲基化: 不敏感
    dcm 甲基化: 不敏感
    CpG甲基化: 阻断酶切

    同裂酶

    SalI

  • 相关产品

    相关产品

    • t1010-monarch-plasmid-miniprep-kit
    • Monarch® DNA 胶回收试剂盒
    • Monarch® PCR & DNA 纯化试剂盒（5 μg）
    • Nuclease-free Water

    单独销售的组分

    • rCutSmart™ 缓冲液
    • 6X 紫色凝胶上样染料

  • 注意事项

    1. CpG 甲基化阻断酶切。

操作说明、说明书 & 用法

  • 操作说明

    1. Optimizing Restriction Endonuclease Reactions
    2. Restriction Digest Protocol
    3. Double Digest Protocol with Standard Restriction Enzymes

  • 使用指南

    • Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
    • Activity of Restriction Enzymes in PCR Buffers
    • Cleavage Close to the End of DNA Fragments
    • Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
    • Double Digests
    • Effects of CpG Methylation on Restriction Enzyme Cleavage
    • Heat Inactivation
    • NEBuffer Activity/Performance Chart with Restriction Enzymes
    • Optimizing Restriction Endonuclease Reactions
    • Restriction Endonucleases – Survival in a Reaction
    • Restriction Enzyme Diluent Buffer Compatibility
    • Restriction Enzyme Tips
    • Single Letter Codes
    • Star Activity
    • Traditional Cloning Quick Guide

工具 & 资源

  • 选择指南

    • Alphabetized List of Recognition Sequences
    • Cleavage of Supercoiled DNA
    • Compatible Cohesive Ends and Generation of New Restriction Sites
    • Dam-Dcm and CpG Methylation
    • Frequencies of Restriction Sites
    • Isoelectric Points (pI) for Restriction Enzymes
    • Isoschizomers
    • NEB Diluent and Buffer Table
    • Recleavable Filled-in 5′ Overhangs
    • Time-Saver™ Qualified Enzymes
    • Why Choose Recombinant Enzymes?

  • Web 工具

    • Competitor Cross-Reference Tool
    • DNA Sequences and Maps Tool
    • Double Digest Finder
    • Enzyme Finder
    • NEBcutter™ v3.0
    • NEBioCalculator®
    • REBASE®

FAQs & 问题解决指南

  • FAQs

    1. Is there any difference in the methylation sensitivity between SalI-HF and SalI?
    2. How does the level of star activity of SalI-HF compare to SalI?
    3. What is the difference between SalI-HF and SalI?
    4. Why does the HF version of the enzyme have a different recommended buffer than the wild type enzyme?
    5. When should I choose the HF version of an enzyme?
    6. When is star activity a concern?
    7. When should I choose the High Fidelity (HF®) version of the enzyme?
    8. Can the change in buffer preference of the HF enzyme be advantageous?
    9. Will the HF enzyme produce elevated star activity when used in a buffer other than the one recommended?
    10. How is the improvement in fidelity of HF restriction endonucleases quantitated?
    11. What is the Fidelity Index (FI)?
    12. What does HF® refer to following the name of a restriction enzyme?
    13. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
    14. Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?
    15. Is Gel Loading Dye, Purple (6X) or Gel Loading Dye, Purple (6X), no SDS compatible with other DNA binding dyes such as SYBR® and GelRed™ during gel electrophoresis?
    16. What does it mean to be Time-Saver™ qualified?
    17. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

  • 问题解决指南

    • Restriction Enzyme Troubleshooting Guide