Express Ni Resin |NEB酶试剂 New England Biolabs

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产品信息

NEBExpress® Ni Resin is an affinity matrix for the isolation and purification of polyhistidine-tagged (His-tagged) fusion proteins. It is intended for use in gravity or pressure flow columns, and batch purifications. NEBExpress Ni Resin is comprised of a highly uniform and stable chemical-tolerant resin, pre-charged with nickel ions on the matrix surface.  It is resistant to a wide range of chemicals, including NaOH, EDTA, DTT and β-Mercaptoethanol.

For larger volume requirements, customized and bulk packaging is available by purchasing through the OEM/Bulks department at NEB. Please contact [email protected] for further information.

Support Matrix: Spherical, agarose based microparticles ranging in size from 10-100 μm.
产品类别:
Amylose Purification (MBP-tag),
NEBExpress MBP Fusion and Purification System,
Bacterial E. coli Protein Expression Products,

Nickel Purification (His-tag),
Affinity Purification Products,
Protein Purification Products,

Protein Expression 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
    • S1428S     4    
        NEBExpress® Ni Resin S1428SVIAL 4 1 x 25 ml 50 %

  • 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

    1 ml of NEBExpress™ Ni Resin will bind ≥ 10 mg of His-tagged fusion protein.

  • 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 Spin Columns

  • 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 (≥ 10 mg/ml) 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 Resin Gravity Flow Typical Protocol (NEB #S1428)
    2. NEBExpress® Ni Resin Batch Binding Typical Protocol (NEB #S1428)
    3. NEBExpress® Ni Resin Pressurized Column Typical Protocol (NEB #S1428)
    4. NEBExpress® Ni Resin Quick Start Protocol (NEB #S1428)
    5. His-tag removal from protein using TEV Protease

工具 & 资源

  • 选择指南

    • Purification Beads, Columns and Resins

FAQs & 问题解决指南

  • FAQs

    1. Can NEBExpress® Ni Resin be used in large scale, pressurized purification systems such as FPLC?
    2. Will extended binding increase the yield of the target protein in the eluate?
    3. Does NEBExpress Ni Resin need to be regenerated with nickel after cleaning?
    4. How can I reduce contaminating proteins in a protein purification when using Ni Resin?
    5. What are the recommendations for pH based elution, as opposed to imidazole?
    6. How can I remove imidazole from a protein sample?
    7. What are to the advantages of performing purification under denaturing conditions?
    8. How can I determine if my target protein remains bound to the resin?
    9. Why is imidazole not necessary in the lysis/binding buffer? Why does the wash buffer contain only 5 mM imidazole?
    10. Why is my protein precipitating on the column?
    11. What gravity column do you recommend?
    12. How can I determine if my target protein remains bound to the resin?

  • 问题解决指南

    • Troubleshooting Guide for Purification using NEBExpress® Ni Resin