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71471 pET Expression System 50b

71471
  
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      Overview

      Replacement Information
      Description
      Overview

      This product has been discontinued.





      pET Expression Systems and pET
      pET Expression Systems and pET Expression Systems plus Competent Cells provide core reagents needed for target gene cloning and expression.

      Components: pET Expression Systems
      Components for pET Expression Systems are similar for all systems unless otherwise stated with the specific pET Expression System description. pET Expression Systems include:
      • 10 µg pET vector DNA (for each indicated plasmid)
      • 0.2 ml BL21 Glycerol Stock
      • 0.2 ml BL21(DE3) Glycerol Stock
      • 0.2 ml BL21(DE3)pLysS Glycerol Stock
      • 0.2 ml Induction Control Glycerol Stock

      Components: pET Expression Systems plus Competent Cells
      pET Expression Systems plus Competent Cells contain all of the components of the specific pET Expression System, as well as the following additional components, unless otherwise stated with the specific pET Expression System description:
      • 0.2 ml NovaBlue Competent Cells
      • 0.2 ml BL21(DE3) Competent Cells
      • 0.2 ml BL21(DE3)pLysS Competent Cells
      • 2 × 0.2 ml SOC Medium
      • 10 µl Test Plasmid

      These components are sufficient for up to 10 transformations in each host.

      Purification and Detection Reagents
      Purification and detection reagents are available separately. For complete product descriptions and ordering information, refer to the Protein Purification and Antibodies, Conjugates & Detection Tools chapters.

      pET Expression System 50b
      The pET-50b(+) vector is designed for cloning and high-level expression of target proteins fused with the 495 aa NusA (Nus•Tag™) protein that is cleavable with the human rhinovirus (HRV) 3C protease (1). The NusA protein has been identified as having the highest potential for solubility when a data base of more than 4000 E. coli proteins was subjected to solubility modeling (2,3). Greater than 85% of the expressed protein was soluble in tests with NusA fusion proteins (2). Cloning sites are also available for producing fusion proteins containing two cleavable N- terminal His•Tag® sequences for detection and purification. The additional N-terminal His•Tag significantly enhances purification yields when utilizing His•Mag™ Agarose Beads for high throughput purification (4). The plasmid contains a strong T7lac promoter, optimized RBS, the coding sequence for the HRV 3C protease cleavage site (LeuGluValLeuPheGlnGlyPro) for N-terminal fusion tag removal and a multiple cloning site that encodes restriction enzyme sites found in many other Novagen expression vectors to facilitate insert transfer. An optional C-terminal S•Tag™ coding sequence is compatible with purification, detection, and quantification (5).




      Commercial use of this Product requires a commercial license from EMD Millipore Corporation. Commercial use shall include but not be limited to (1) use of the Product or its components in manufacturing; (2) use of the Product or its components to provide a service, information, or data to others in exchange for consideration; (3) use of the Product or its components (or any derivatives thereof) for therapeutic, diagnostic or prophylactic purposes (including as part of a device, chip, assay or other product); or (4) resale of the Product or its components, whether or not such Product or its components are resold for use in research. Nothing contained herein shall be deemed to represent or warrant that additional third party rights are not required for use of this Product.
      Catalogue Number71471
      Brand Family Novagen®
      References
      References1. Walker, P.A., Leong L. E.-C., Ng, P.W.P., Tan, S.H.,Waller, S., Murphy, D. and Porter, A.G. (1994) Bio/Technology 12, 601-605.

      2. Harrison, R.G. (2000) inNovations 11, 4-7.

      3. Davis, G.D., Elisee, C., Newham, D.M. and Harrison, R.G. (1998) Biotechnol. Bioeng. 65, 382-388.

      4. Novy, R. and Drott, D. (2002) inNovations 14, 12-13.

      5. Kim, J.S. and Raines, R.T. (1993) Protein Science 2, 348-356.
      Product Information
      10 µgpET-50b(+) DNA
      0.2 mlBL21 Glycerol Stock
      0.2 mlBL21(DE3) Glycerol Stock
      0.2 mlBL21(DE3)pLysS Glycerol Stock
      0.2 mlInduction Control Glycerol Stock
      Fusion tagHis•Tag, Nus•Tag, S•Tag
      Applications
      Biological Information
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Shipped with Blue Ice or with Dry Ice
      Toxicity Standard Handling
      Storage ≤ -70°C
      Do not freeze Ok to freeze
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      71471 0

      Documentation

      pET Expression System 50b Certificates of Analysis

      TitleLot Number
      71471

      References

      Reference overview
      1. Walker, P.A., Leong L. E.-C., Ng, P.W.P., Tan, S.H.,Waller, S., Murphy, D. and Porter, A.G. (1994) Bio/Technology 12, 601-605.

      2. Harrison, R.G. (2000) inNovations 11, 4-7.

      3. Davis, G.D., Elisee, C., Newham, D.M. and Harrison, R.G. (1998) Biotechnol. Bioeng. 65, 382-388.

      4. Novy, R. and Drott, D. (2002) inNovations 14, 12-13.

      5. Kim, J.S. and Raines, R.T. (1993) Protein Science 2, 348-356.

      User Protocols

      Title
      TB053 Academic and Non-profit Laboratory Assurance Letter
      TB055 pET System Manual

      Vector Map

      Title
      TB418VM pET-50b(+) Vector Map

      Vector Sequence

      Title
      pET-50b(+) Vector Sequence