69954 pET-31b(+) DNA, AlwN I Digest - Novagen

69954
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      Description
      Overview

      This product has been discontinued.





      This kit is available as a convenient supplement to the pET Peptide Expression System 31b, which is designed for expression and purification of target peptides (1). In pET-31b(+), peptide coding sequences are placed downstream of a 125 amino acid (aa) ketosteroid isomerase (KSI) gene and upstream of a His•Tag® sequence. Use of the unique AlwN I cloning site enables unidirectional insertion of coding sequences immediately adjacent to a methionine residue. For high-yield production, small (10–25 aa) peptide coding sequences can be cloned as tandem repeats interspersed with single methionine residues. The KSI-peptide-His•Tag fusion protein is expressed at high levels in the presence of T7 RNA polymerase provided by the recA, protease deficient BLR(DE3)pLysS host. The fusion protein is purified in one step under denaturing conditions using one of Novagen's His•Bind® metal chelation resins. It is then cleaved with CNBr to release the His•Tag peptide, insoluble KSI, and monomer peptide units each terminating with a homoserine lactone. The purified peptide can be easily converted to an amide or homoserine, or reacted with other compounds, such as analogs of fluorescein and biotin, to produce C-terminal derivatized peptides. The system has been used to make biologically active peptides of 14–67 aa at yields of > 50 mg purified peptide per liter of culture (1).

      The AlwN I digested, dephosphorylated vector is ready to accept appropriate inserts containing ...ATG-3′ and 3′-TAC... overhangs on the top and bottom strands, respectively. A Control Insert is provided to verify performance and ensure optimal cloning efficiency.

      Features
      • High yield expression of peptides and small proteins
      • His•Tag sequence facilitates protein purification
      • Prepared vector facilitates cloning with minimal background
      • Special host strain inhibits degradation of expressed proteins..






      This product is sold for internal research use only. Any commercial use of this product, its components, and/or any derivatives thereof (including but not limited to proteins produced using the product or its components) (together and hereinafter the 'EMD Product') requires signature of a written commercial use agreement with EMD Millipore Corporation or its successor-in-interest. Commercial use shall include but not be limited to: (1) use of the EMD Product to manufacture products for sale to third parties; (2) use of the EMD Product to provide services, information, or data to third parties in exchange for consideration; (3) use of the EMD Product for therapeutic, diagnostic or prophylactic purposes (including as part of a device, chip, assay or other product); or (4) resale of the EMD Product, whether or not such EMD Product is resold for research use. Nothing contained herein shall be deemed to represent or warrant that additional third party rights are not required for use of the EMD Product. Please direct any questions on these use restrictions to: licensing@milliporesigma.com.

      Catalogue Number69954
      Brand Family Novagen®
      References
      References1. Kuliopulos, A. and Walsh, C.T. (1994) J. Am. Chem. Soc. 116, 4599–4607.
      Product Information
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      Quality LevelMQ100
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      Ship Code Shipped with Blue Ice or with Dry Ice
      Toxicity Standard Handling
      Storage ≤ -70°C
      Avoid freeze/thaw Avoid freeze/thaw
      Do not freeze Ok to freeze
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      Documentation

      pET-31b(+) DNA, AlwN I Digest - Novagen Certificates of Analysis

      TitleLot Number
      69954

      References

      Reference overview
      1. Kuliopulos, A. and Walsh, C.T. (1994) J. Am. Chem. Soc. 116, 4599–4607.

      Citations

      Title
    • Min-Ju Kim, et al. (2007) NMR structural studies of interactions of a small, nonpeptidyl Tpo mimic with the thrombopoietin receptor extracellular juxtamembrane and transmembrane domains. Journal of Biological Chemistry 282, 14253-14261.
    • User Protocols

      Title
      TB053 Academic and Non-profit Laboratory Assurance Letter
      TB055 pET System Manual
      TB104 pET Peptide Expression System 31

      Vector Map

      Title
      TB098VM pET-31b(+) Vector Map