NEBExpress^® Ni-NTA Magnetic Beads

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
S1423S     4       NEBExpress® Ni-NTA Magnetic Beads S1423SVIAL 4 1 x 1 ml 10 %   2X IMAC Buffer B1076SVIAL 4 1 x 30 ml 2 X   2M Imidazole B1077SVIAL 4 1 x 2 ml 2 M
S1423L     4       NEBExpress® Ni-NTA Magnetic Beads S1423SVIAL 4 5 x 1 ml 10 %   2X IMAC Buffer B1076SVIAL 4 5 x 30 ml 2 X   2M Imidazole B1077LVIAL 4 1 x 10 ml 2 M

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 and detergents.
  
  
• High specific binding of His-tagged proteins from various expression systems with purities of >95%.
  
  
• NTA securely coordinates metal ions through four coordination sites which results in low nickel ion leaching.
  
  
• Can be used with common cell lysis reagents and a variety of buffer additives.

Companion Products

• TEV Protease
• 6-Tube Magnetic Separation Rack
• 12-Tube Magnetic Separation Rack
• 96-Well Microtiter Plate Magnetic Separation Rack
• 50 ml Magnetic Separation Rack

1. Do not freeze.
2. Do not store in buffer containing imidazole.
3. 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-NTA 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).
4. Protein yield and purity are dependent upon the expression level, conformation and solubility characteristics of the recombinant fusion protein. For best results, perform a small-scale test to estimate the expression level and determine the solubility of each His-tagged protein.
5. Avoid using protease inhibitors or other additives that contain chelators, such as EDTA, or strong reducing agents, such as DTT or β-mercaptoethanol, which will impact the performance of the NEBExpress™ Ni-NTA Magnetic Beads.
6. 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).
7. NEBExpress™ Ni-NTA Magnetic Beads yield highly variable binding capacities dependent on the target and conditions. The binding capacity value (≥ 7.5 mg/ml) for this product was determined by performing a mock purification from crude cell lysates supplemented with his-tagged target protein. The reported capacities of the most commercial IMAC sorbents are up to 40 mg/mL. However, these values refer to isolated pure proteins or synthetic mixtures, while capacities for isolation of the target protein from complex sources are typically lower.

1. Bolanos-Garcia et al (2006). BBA. 1760, 1304-1313.
2. Samuelson, J (2016). Purifying Recombinant His-Tagged Proteins. Genetic Engineering & Biotechnology News. 26-27.

1. NEBExpress^® Ni-NTA Magnetic Beads Typical Reaction Protocol
2. Quick Start Protocol for NEBExpress^® Ni-NTA Magnetic Beads
3. His-tag removal from protein using TEV Protease

• NEBExpress^® Cell-free E. coli Protein Synthesis System
• Automated Cell-free Protein Expression and Purification for High-Throughput Screening using NEBExpress^® Cell-free E. coli Protein Synthesis System and NEBExpress Ni-NTA Magnetic Beads

• Purification Beads, Columns and Resins

1. Which E. coli strain do you recommend to reduce contaminating proteins?
2. The recovery of the His-tagged target protein is low relative to contaminant proteins. Should I add more NEBExpress^® Ni-NTA Magnetic Beads to improve recovery of my target protein?
3. How can I reduce contaminating proteins in my Ni-NTA His-tag protein purification?
4. My purification failed and/or why is my yield lower than expected?
5. How can I remove imidazole from a protein sample?
6. What is the minimum elution volume that should be used?
7. Why is there is so much target protein in the column flow through?
8. Why is there variability in my high throughput assay?
9. Should the crude extract be diluted with lysis buffer to increase the volume for the binding step?
10. What is a typical protocol for lysing cells?
11. What are the recommendations for pH based elution, as opposed to imidazole?
12. What are to the advantages of performing purification under denaturing conditions?

• Protein Expression & Purification Brochure
• Purification Beads, Columns and Resins Brochure

• Purifying Recombinant His-Tagged Proteins
• Over 40 years in protein expression and purification – a historical perspective

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.

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]

• S1423S_L_v1

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]

• S1423L_v1_10043363
• S1423S_v1_10043361
• S1423S_v1_10056435
• S1423S_v1_10065736
• S1423S_v1_10065935
• S1423S_v1_10067059
• S1423S_v1_10067691
• S1423S_v1_10068231
• S1423S_v1_10070717
• S1423L_v1_10080721
• S1423L_v1_10096772
• S1423S_v1_10111873
• S1423L_v1_10111872
• S1423L_v1_10115390
• S1423L_v1_10118373
• S1423S_v1_10118374
• S1423L_v1_10128992
• S1423S_v1_10128993
• S1423S_v1_10140252
• S1423S_v1_10144361
• S1423L_v1_10140253
• S1423L_v1_10153423
• S1423L_v1_10157793
• S1423S_v1_10153424
• S1423S_v1_10164693
• S1423L_v1_10178163
• S1423S_v1_10176101
• S1423S_v1_10180168
• S1423S_v1_10183763
• S1423S_v1_10195875
• S1423L_v1_10208886
• S1423S_v1_10206254
• S1423L_v1_10217670
• S1423S_v1_10218938
• S1423L_v1_10232698
• S1423S_v1_10226292
• S1423L_v1_10239130
• S1423S_v1_10239131
• S1423L_v1_10249055
• S1423S_v1_10247094
• S1423S_v1_10250779
• S1423L_v1_10257216
• S1423S_v1_10256976

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

• NEBExpress^® Ni-NTA Magnetic Beads
• 2X IMAC Buffer
• 2M Imidazole

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.