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Home Discontinued Products CoA-SH

CoA-SH

  • Catalog # S9352 was discontinued on October 03, 2014 ;
  • Product Information
    CoA-SH is a reactive building block for the one-step synthesis of ACP-tag or MCP-tag substrates from maleimide-containing precursors including fluorophores, peptides or oligonucleotides. It is suitable for linkage to maleimides and iodoacetamides.

    The ACP-tag and MCP-tag are polypeptide tags (8 kD) based on the acyl carrier protein. MCP-tag contains two mutations (D36T and D39G). Both allow the specific, covalent attachment of virtually any molecule to a protein of interest. Substrates are derivates of Coenzyme A (CoA). In the labeling reaction, the substituted phosphopantetheine group of CoA is covalently attached to a conserved serine residue of the ACP-tag or the MCP-tag by a phosphopantetheine transferase (SFP Synthase or ACP Synthase).

    While the ACP Synthase (NEB #P9301) will label predominantly the ACP-tag, the SFP Synthase (NEB #P9302) will modify ACP-tag and MCP-tag. CoA substrates react with both the ACP-tag and MCP-tag.

    Having no cysteines, the ACP-tag and the MCP-tag are particularly suited for specifically labeling cell-surface proteins, and should be useful for labeling secreted proteins with disulfide bridges such as antibodies.

    There are two steps to using this system: sub-cloning and expression of the protein of interest as an ACP-tag or MCP-tag fusion, and labeling of the fusion protein using the appropriate synthase with the CoA substrate of choice. In this document, the labeling of the fusion proteins with CoA substrates is described. The cloning of ACP-tag and MCP-tag protein fusions is described in the documentation supplied with the ACP-tag or MCP-tag plasmids.

    Figure 1. Structure of CoA-SH (MW 785.3 g/mol)
    Product Categories:
    Discontinued Products
    • Properties & Usage

      Reaction Conditions

      Reaction Conditions for Chemical Coupling:
      CoA-SH is a maleimide-reactive building block which can be used for the one-step synthesis of CoA substrates from maleimide-containing precursors. This building block allows you to make custom CoA substrates for labeling ACP-tag or MCP-tag fusion proteins for a wide range of applications. 

      A typical reaction is performed at 30°C overnight by using N,N-dimethylformamide (DMF) and Tris-buffer (pH 7.5) as solvents. Equimolar quantities of CoA-SH and reactive maleimide are recommended.

    • Product Notes
      1. Storage:
        Store the vials containing the CoA substrate at -20°C. The CoA-SH building block should be dissolved in Tris buffer pH 7.5. Choose a solvent volume that will be compatible with the final use. Unused building block in Tris buffer pH 7.5 should be stored at -20°C under argon.
    • References
      1. Yin, J. et al. (2005). Chemistry & Biology. 12, 999-1006.
      2. George, N. et al. (2004). JACS. 126, 8896-8897.
      3. Vivero-Pol, L. et al. (2005). JACS. 127, 12770-12771.
  • Protocols, Manuals & Usage
  • Tools & Resources
  • FAQs & Troubleshooting
  • Quality, Safety & Legal
    • Quality Control Assays
      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.
    • Safety Data Sheets
      The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
    • Datacards
      The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.)
    • Legal and Disclaimers
      This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

      While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

      For more information about commercial rights, please contact NEB's Global Business Development team at gbd@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.

      Licenses

      Cellular Imaging and Analysis (i.e., SNAP and CLIP products)

      Notice to Buyer/User: The Buyer/User has a non-exclusive license to use this system or any component thereof for RESEARCH AND DEVELOPMENT ONLY. Commercial use of this system or any components thereof requires a license from New England Biolabs, Inc., 240 County Road, Ipswich, MA 01938. For detailed information see our Terms of Use .

      The products and/or their use may be covered by one or more of the following patents and patent applications:
      7,939,284 (Methods for Using O6-Alkylguanine-DNA-Alkyltransferases)
      7,888,090 (Mutants of O6-Alkylguanine-DNA-Alkyltransferases)
      8,163,479 Specific Substrates for  O6-Alkylguanine-DNA-Alkyltransferases)
      8,178,314 (Pyrimidines Reacting With O6-Alkylguanine-DNA-Alkyltransferases)
      PCT/EP2007/057597 (Labeling of Fusion Proteins with Synthetic Probes)
      EP07117800 (Drug Delivery)
      EP07117802 (Drug Delivery)
      EP07120288 (GTPase-Transient Protein Protein Interactions)
      These patents and patent applications are owned by Covalys, or owned by the Ecole Polytechnique Fédérale de Lausanne (EPFL) and exclusively licensed to Covalys and NEB.

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