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NEBExpress® Ni-NTA Magnetic Beads

Catalog # Concentration Size List Price Quantity Your Price
S1423L 10 % 5 ml $1,250.00
$1,125.00
S1423S 10 % 1 ml $284.00
$255.60
Catalog # Size List Price Your Price
S1423L 5 ml $1,250.00
$1,125.00
S1423S 1 ml $284.00
$255.60
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
  • An affinity matrix for the small-scale isolation and purification of polyhistidine-tagged (His-tagged) fusion proteins in manual or automated formats.
  • Prepared with agarose based, super-paramagnetic microparticles which provide high binding capacity and fast magnetic response permitting high throughput and scalable purification strategies of His tagged-fusion proteins.
  • NTA coordination exhibits low Nickel ion leaching.
  • Tolerates a wide range of conditions, including the presence of protein denaturants and detergents. Compatible with commercially available detergent based cell lysis reagents.
  • Elution of the target protein may be achieved by protonation, ligand exchange (with imidazole) or extraction of the metal ion by a stronger chelator (e.g. EDTA).
  • Extremely low non-specific binding properties permit immobilized fusion proteins to be used in subsequent experiments to capture or pull down interacting proteins from crude cell lysates
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    Introduction to the NEBExpress® Cell-free Protein Synthesis System

An affinity matrix for the small-scale isolation and purification of polyhistidine-tagged (His-tagged) fusion proteins in manual or automated formats. Immobilized Metal Affinity Chromatography (IMAC) purifications employing Ni-NTA (nickel-nitrilotriacetic acid) magnetic beads can be performed under native or denaturing conditions which permit efficient binding and purification of insoluble proteins, proteins that aggregate in inclusion bodies, or proteins that possess a tertiary structure that sequester the polyhistidine affinity tag. Immobilized His-tagged proteins can be used in subsequent experiments to pull-down proteins that may interact with the immobilized protein.

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.

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  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
  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.
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.

Companion Products
Notes
  • Do not freeze.
  • Do not store in buffer containing imidazole.
  • 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).
  • 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.
  • 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.
  • 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).
  • 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.
References
  • Bolanos-Garcia et al (2006). BBA. 1760, 1304-1313.
  • Samuelson, J (2016). Purifying Recombinant His-Tagged Proteins. Genetic Engineering & Biotechnology News. 26-27.
Quality Control Assay
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 Data Sheets
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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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