Product Class: Other

NEBExpress® Ni Resin

NEBExpress® Ni Resin is also available in spin column format, NEB #S1427


Catalog #S1428

Product Introduction

 

  • Purification of ≥10 mg His-tagged protein per 1 ml of resin
  • Intended for use in gravity or pressure flow columns, and batch purifications
  • High specific binding of His-tagged proteins yielding purities of >95%
  • Strong nickel ion binding provides excellent resistance to EDTA and reducing agents. Compatible with commercially available detergent-based cell lysis reagents
  • Isolation and purification of His-tagged fusion proteins under native or denaturing conditions
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Product Information

Description

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 custom@neb.com for further information.

Support Matrix: Spherical, agarose based microparticles ranging in size from 10-100 μm.
This product is related to the following categories:
Amylose Purification (MBP-tag),
NEBExpress MBP Fusion and Purification System,
Nickel Purification (His-tag),
Bacterial E. coli Protein Expression Products,
Affinity Purification Products,
Protein Purification Products,
Protein Expression Products,
This product can be used in the following applications:
MBP Affinity Tag,
Fusion Protein Cleavage,
Pull Down Assays,
His-tagged Protein Expression & Purification,
Affinity Purification & Expression Tags,
Protein Purification,
Protein Analysis Tools

Properties & Usage

Storage Buffer

20% Ethanol

Support Matrix

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

Binding Capacity

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

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

Product Notes

  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.

References

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

Protocols, Manuals & Usage

Protocols

  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

Tools & Resources

Selection Charts

FAQs & Troubleshooting

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?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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