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
Improved purity of target proteins isolated by IMAC
Application Features
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
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.
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Specifications & Change Notifications
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]
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]
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Licenses
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.
