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NiCo21(DE3) Competent E. coli

Catalog # Concentration Size List Price Quantity Your Price
C2529H 20 x 0.05 ml $628.00
$565.20
Catalog # Size List Price Your Price
C2529H 20 x 0.05 ml $628.00
$565.20
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Chemically competent E. coli cells suitable for T7 expression with improved purity of target proteins isolated by IMAC.

  • Superior alternative to BL21(DE3) for routine protein expression
  • Deficient in proteases Lon and OmpT
  • Resistant to phage T1 (fhuA2)
  • Free of animal products
  • No dry ice surcharge on competent cell shipments
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    Fast transformation plating with glass beads
Chemically competent E. coli cells derived from BL21(DE3). Poly-histidine tagged recombinant proteins that are isolated by immobilized metal affinity chromatography (IMAC) are often contaminated with significant amounts of endogenous E. coli metal binding proteins. The protein expression strain NiCo21(DE3) has been engineered to minimize E. coli protein contamination of IMAC fractions: GlmS is mutated to eliminate binding to IMAC resins and three other proteins (SlyD, ArnA and Can) are tagged to enable rapid removal by chitin affinity chromatography.
Highlights
  • Superior alternative to BL21(DE3) for routine protein expression
  • Improved purity of target proteins isolated by IMAC
  • Identical growth characteristics as BL21(DE3)
  • Deficient in proteases Lon and OmpT
  • Resistant to phage T1 (fhuA2)
  • Free of animal products
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  pUC19 Vector N3041AVIAL -20 1 x 0.025 ml 50 pg/µl
  SOC Outgrowth Medium B9020SVIAL 4 1 x 25 ml
  NiCo21(DE3) Competent E. coli C2529HVIAL -80 20 x 0.05 ml
Features
  • Improved purity of target proteins isolated by IMAC
Application Features

Improved purity of His-tagged proteins with NiCo21(DE3) Improved purity of His-tagged proteins with NiCo21(DE3)
A) Expression of Glutamyl tRNA Synthetase (6-His) in NiCo21(DE3) Competent E. coli followed by Ni-NTA purification. Eluent (E) from a Ni-NTA column was passed over a chitin column. The protein of interest elutes in the flow through (FT), while the CBD-tagged metal binding proteins remain bound (B) to the chitin resin (NEB #S6651 ). Purifications were performed according to manufacturers' recommended conditions. B) Contaminants ArnA, SlyD and Can are confirmed by Western blot using Anti-CBD Antibody (NEB #E8034 ).

Two-step purification of target protein that has been expressed in the NiCo21(DE3) strain of E. coli
process

Properties & Usage

Antibiotic for Plasmid Selection

Antibiotics for Plasmid Selection Working Concentration
Ampicillin 100 µg/ml
Carbenicillin 100 µg/ml
Chloramphenicol 33 µg/ml
Kanamycin 30 µg/ml
Streptomycin 25 µg/ml
Tetracycline 15 µg/ml

Shipping Notes

  • Ships on dry ice


Companion Products
Materials Sold Separately
Notes
  • CAUTION: This product contains DMSO, a hazardous material. Review the MSDS before handling.
  • STORAGE AND HANDLING: Competent cells should be stored at -80°C. Storage at -20°C will result in a significant decrease in transformation efficiency. Cells lose efficiency whenever they are warmed above -80°C, even if they do not thaw.
References
  • Bolanos-Garcia and Davies (2006). Biochimica et Biophysica Acta. 1760, 1304-1313.
  • Carine Robichon, Jianying Luo, Thomas B. Causey, Jack S. Benner, and James C. Samuelson (2011). Applied and Environmental Microbiology. 77, 4634-4646.
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
Legal And Disclaimer

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.Information for Non-U.S. Customers
For European Customers
The NiCo21(DE3) E.coli strain is genetically modified to carry the can::CBD, arnA::CBD and slyD::CBD gene fusions. The CBD (chitin binding domain) DNA sequence originates from Bacillus circulans. As a condition of sale, this product must be in accordance with all applicable local legislation and guidelines including EC Directive 90/219/EEC on the contained use of genetically modified organisms.

For All Other Non-U.S. Customers
The parental strain of NiCo21(DE3) E. coli is BL21(DE3) (derivative of E.coli B, Delbrück and Luria, 1942). Although the parental strain is generally classified as Biosafety Level 1 (BL-1), we recommend that you consult the safety department of your institution to verify the Biosafety Level.

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