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Express Ni Spin Columns |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

Spin columns containing an affinity matrix for the small-scale isolation and purification of polyhistidine-tagged (His-tagged) fusion proteins. Immobilized Metal Affinity Chromatography (IMAC) purifications employing NEBExpress® Ni Spin columns can be performed under native or denaturing conditions, yielding highly pure target in a single protein purification step.  This enables screening of expression conditions and streamlines the functional and structural characterization of the target protein.

Support Matrix: Spherical, agarose based microparticles ranging in size from 10-100 μm.
 

Figure 1: NEBExpress Ni Spin Column Quick Start Protocol

Express Ni Spin Columns |

Figure 2: Purification of His-tagged fusion proteins using NEBExpress Ni Spin Columns

Express Ni Spin Columns |

NEBExpress Ni Spin Columns were equilibrated and loaded with 500 μl of crude extract from E. coli containing a plasmid that expresses either His6-GluRS or His6-NDK fusion protein. Columns were washed, eluted and analyzed by SDS-PAGE to confirm low non-specific binding of extract proteins and high iso­lation of target. Marker (M) contains Color Prestained Protein Standard (NEB #P7719), Lane 1: GluRS protein load, Lane 2: GluRS flow through, Lane 3: GluRS 1st elution, Lane 4: GluRS 2nd elution, Lane 5: NDK protein Load, Lane 6: NDK flow through, Lane 7: NDK 1st elution, Lane 8: NDK 2nd elution.
产品类别:
Amylose Purification (MBP-tag),
Nickel Purification (His-tag),
Affinity Purification Products,

Protein Purification Products

应用:
MBP Affinity Tag,
Fusion Protein Cleavage,
Pull Down Assays,

His-tagged Protein Expression & Purification,
Affinity Purification & Expression Tags,
Protein Purification,

Protein Analysis Tools

  • 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
    • S1427S     4    
        NEBExpress® Ni Spin Columns S1427SVIAL 4 10 x 1 Each
      (0.1 ml)       		Not Applicable
        2X IMAC Buffer B1076SVIAL 4 1 x 30 ml 2 X
        2M Imidazole B1077SVIAL 4 1 x 2 ml 2 M
    • S1427L     4    
        NEBExpress® Ni Spin Columns S1427SVIAL 4 25 x 1 Each
      (0.1 ml)       		Not Applicable
        2X IMAC Buffer B1076SVIAL 4 2 x 30 ml 2 X
        2M Imidazole B1077LVIAL 4 1 x 10 ml 2 M

  • 特性和用法

    贮存溶液

    20% Ethanol

    支持介质

    Spherical, agarose based microparticles ranging in size from 10-100 μm.

    结合容量

    Varies with target, ≥ 1 mg His-tagged fusion protein per column.

  • 优势和特性

    Features

    • Isolation and purification of His-tagged fusion proteins under native or denaturing conditions as IMAC tolerates a wide range of conditions, including the presence of protein denaturants, reducing agents and detergents.
    • High specific binding of His-tagged proteins from various expression systems yielding milligram quantities of target protein with purities of >95%.
    • Can be used with common cell lysis reagents and a variety of buffer additives

  • 相关产品

    相关产品

    • NEBExpress® Ni-NTA 磁珠
    • TEV Protease
    • Amylose Resin
    • Amylose Resin High Flow
    • 几丁质树脂
    • NEBExpress® Ni Resin

  • 注意事项

    1. Do not freeze.
    2. Several E. coli proteins are capable of binding immobilized metal affinity matrices with moderate to high affinity. The most common contaminants include SlyD (28kDa), GlmS (67kDa), ArnA (74kDa) and carbonic anhydrase (25 kDa) (1). The named contaminants may be eliminated from the Ni column elution fraction by employing NiCo21(DE3) (NEB# C2529H) as the expression host and then performing a second binding step with chitin magnetic beads (NEB# E8036) (2). 
    3. Protein yield and purity are dependent upon the expression level, conformation and solubility characteristics of the recombinant fusion protein.  For best results, estimate the expression level of the His-tagged protein of interest by first running a sample of the crude lysate on an SDS-PAGE gel. 
    4. Unlike most IMAC resins, the unique chemistry employed in NEBExpress™ Ni Resin provides chemical tolerance to chelating agents such as EDTA and reducing agents such as DTT.  Please consult the chemical compatibility table for more information. 
    5. Polyhistidine tags do not typically compromise the biological protein function and are not considered immunogenic, however if cleavage of the His-tag is necessary we recommend using TEV Protease (NEB# P8112).
    6. Ni resin yields highly variable binding capacities dependent on the target and conditions. The binding capacity value (≥ 1 mg/column) for this product was determined by performing a mock purification from crude cell lysates expressing a His-tagged target protein. 

  • 参考文献

    1. Bolanos-Garcia et al (2006). BBA. 1760, 1304-1313.
    2. Samuelson, J (2016). GEN. 26-27.

操作说明、说明书 & 用法

  • 操作说明

    1. NEBExpress® Ni Spin Columns Quick Start Protocol (NEB #S1427)
    2. NEBExpress® Ni Spin Column Reaction Protocol (NEB #S1427)
    3. His-tag removal from protein using TEV Protease

  • 应用实例

    • NEBExpress® Cell-free E. coli Protein Synthesis System

工具 & 资源

  • 选择指南

    • Purification Beads, Columns and Resins

FAQs & 问题解决指南

  • FAQs

    1. How much lysate can be loaded onto a single NEBExpress Ni Spin Column?
    2. Will extended binding increase the yield of the target protein in the eluate?
    3. What is the minimum recommended load volume?
    4. Is it necessary to cap the column during each centrifugation step?
    5. Why is the liquid not completely removed during the centrifugation step?
    6. I’ve prepared my lysate with too much buffer and the target is very dilute, can I apply more lysate by repeating the load steps?
    7. How can I reduce contaminating proteins in a Ni spin column protein purification?
    8. What are the recommendations for pH based elution, as opposed to imidazole?
    9. How can I remove imidazole from a protein sample?
    10. What are to the advantages of performing purification under denaturing conditions?
    11. How can I determine if my target protein remains bound to the resin?
    12. Why is imidazole not necessary in the lysis/binding buffer? Why does the wash buffer contain only 5mM imidazole?

  • 问题解决指南

    • Troubleshooting Guide for Purification using NEBExpress® Ni Spin Columns

Monarch® RNA Cleanup Columns (10 μg) |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

The Monarch RNA Cleanup Columns (10 µg) can be used to purify up to 10 µg of nucleic acid from enzymatic reactions. The columns are designed without the use of a retaining ring, helping to ensure no buffer retention and no carryover of contaminants. Nucleic acid can be eluted in as little as 6 μl and is ready for use in a variety of downstream applications including RT-PCR, RNA library prep for NGS, and labelling.

Monarch RNA Cleanup Columns are also available for 50 µg (NEB #T2047) and 500 µg (NEB #T2057) binding capacities.

Properties

Also included as a component in: T2030 Monarch RNA Cleanup Kit (10 µg)
Product format: Spin columns and collection tubes for nucleic acid purification
Technology and purification format: Silica membrane-based purification technology
Provided as: 100 columns and 100 collection tubes
Suitable for: Cleaning up nucleic acid from a variety of enzymatic reactions; Concentration of nucleic acid into small volumes

 

产品类别:
RNA Cleanup Products,
RNA Extraction and Purification,
Nucleic Acid Purification Products

  • 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
    • T2037L     25    
        Monarch® RNA Cleanup Columns (10 μg) T2037-1 25 2 x 50 columns Not Applicable
        Monarch® Collection Tubes II T2018-1 25 2 x 50 tubes Not Applicable

操作说明、说明书 & 用法

  • 使用指南

    • Avoiding Ribonuclease Contamination
    • Guidelines for RNA Quantitation
    • Guidelines for Working with RNA During RNA Cleanup

  • 应用实例

    • A Practical Guide to Analyzing Nucleic Acid Concentration and Purity with Microvolume Spectrophotometers

FAQs & 问题解决指南

  • FAQs

    1. What is the maximum binding capacity of the Monarch RNA Cleanup Column provided with the Monarch RNA Cleanup Kit?
    2. What is the smallest volume of nuclease-free water that can be used for elution with the Monarch RNA Cleanup Columns?
    3. Can I get better recovery with the Monarch RNA Cleanup Kits if I do a second elution with my eluent from the first elution?
    4. What factors affect my (A260/A230) when using the Monarch RNA Cleanup Kits?
    5. What size RNA can be purified with the Monarch RNA Cleanup Kit?
    6. Can I use the Monarch RNA Cleanup Kit to cleanup up my DNase I-treated RNA?
    7. Can I use the Monarch RNA Cleanup Kits to purify RNA from agarose gels?
    8. Can I use the Monarch RNA Cleanup Kits to cleanup RNA after a TRIzol®/chloroform extraction?
    9. Are the Monarch RNA Cleanup Kits (NEB # T2030, #T2040, #T2050) compatible with Luna RT-qPCR reagents?
    10. Are the Monarch RNA Cleanup Kits (NEB #T2030, #T2040, #T2050) compatible with NEBNext reagents for RNA library prep?
    11. How can I assess RNA integrity and purity?
    12. Are the columns in the Monarch RNA Cleanup Kits the same as those in the Monarch Total RNA Miniprep Kit (NEB #T2010)?
    13. Can I purchase Monarch buffers and columns separately?
    14. Can I do an on-column DNase I treatment with the Monarch RNA Cleanup Columns?
    15. Can the Monarch RNA Cleanup Kits (NEB #T2030, #T2040, #T2050) also be used to purify DNA?
    16. Can the Monarch RNA Cleanup Kits be used for RNA extraction?

  • 问题解决指南

    • Troubleshooting Guide for RNA Cleanup

Monarch® Plasmid Miniprep Columns |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

Monarch Plasmid Miniprep Columns, supplied with the Monarch Plasmid Miniprep Kit, have been custom designed to deliver excellent performance for your plasmid purification. Monarch Columns are designed without a frit (which is commonly used in purification columns to hold the membrane in place), eliminating buffer retention and the risk of carry-over contamination. The tapered design of the Miniprep Column enables elution in as little as 30 μl. Monarch columns use less plastic than conventional columns, reducing their environmental footprint without compromising performance. The columns fit snugly into the collection tubes to enable easy handling. The Monarch Miniprep Columns also feature a convenient tab so that you can label your columns easily without removing them from your tube rack. Monarch columns are suitable for use with centrifugation and vacuum purification protocols, and are supplied with collection tubes.

Monarch Plasmid Miniprep Column Design
Monarch® Plasmid Miniprep Columns |

重点

  • No buffer retention/risk of carryover contamination
  • Low elution volume (≥ 30 μl)
  • Convenient tab for easy labeling
  • Snug fit into collection tubes for easy handling
  • Made with less plastic than conventional columns
产品类别:
Plasmid Purification Products,
Nucleic Acid Purification Products

应用:
Nucleic Acid Purification

  • 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
    • T1017L     25    
        Monarch® Plasmid Miniprep Columns T1017-2 25 2 x 50 columns Not Applicable

  • 特性和用法

    结合容量

    20 µg

  • 相关产品

    相关产品

    • t1010-monarch-plasmid-miniprep-kit
    • Monarch® 质粒重悬缓冲液（B1）
    • Monarch® 质粒裂解液（B2）
    • Monarch® 质粒中和缓冲液（B3）
    • Monarch® 质粒洗涤缓冲液 1
    • Monarch® 质粒洗涤缓冲液 2
    • Monarch® DNA 洗脱缓冲液
    • Monarch® DNA 纯化离心柱（5 μg）
    • Monarch® DNA 胶回收试剂盒
    • Monarch® PCR & DNA 纯化试剂盒（5 μg）
    • Quick-Load® 紫色 1 Kb Plus DNA Ladder
    • 1 kb Plus DNA Ladder

    单独销售的组分

    • Monarch® 质粒重悬缓冲液（B1）
    • Monarch® 质粒裂解液（B2）
    • Monarch® 质粒中和缓冲液（B3）
    • Monarch® 质粒洗涤缓冲液 1
    • Monarch® 质粒洗涤缓冲液 2
    • Monarch® DNA 洗脱缓冲液

操作说明、说明书 & 用法

  • 操作说明

    1. Monarch® Plasmid DNA Miniprep Kit Protocol (NEB #T1010)

  • 使用指南

    • Back to basics: Important things to keep in mind when purifying plasmids and DNA fragments
    • Dos and Don’ts of Plasmid Minipreps
    • Usage Guidelines for the Monarch Plasmid Miniprep Kit (#T1010) When Working with Low Copy Plasmids

  • 应用实例

    • Using Exonuclease V (RecBCD) to Eliminate Residual Genomic DNA When Purifying Low Copy Plasmids with the Monarch® Plasmid Miniprep Kit
    • A Practical Guide to Analyzing Nucleic Acid Concentration and Purity with Microvolume Spectrophotometers

FAQs & 问题解决指南

  • FAQs

    1. Are plasmids recovered using the Monarch Plasmid Miniprep Kit endotoxin free?
    2. I have used 5 ml of cell culture for plasmid isolation with the Monarch Plasmid Miniprep kit and I am obtaining low amounts of plasmid and/or contaminating gDNA. Why is this, and what are your suggestions to improve yield and purity?
    3. If I need to clean up more than 5 μg of DNA, is there another higher capacity column alternative to the columns included in the Monarch PCR & DNA Cleanup kit (NEB #T1030)?

  • 问题解决指南

    • Troubleshooting Guide for DNA Cleanup and Plasmid Purification