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  • pSNAPf-Cox8A Control Plasmid

    Discontinued Date

    01/01/2013
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    Categories:
    Discontinued Products

    Description

    This control plasmid contains the gene encoding the Cytochrome C oxidase, subunit 8-2 (COX8-2) protein cloned upstream of the SNAPf coding sequence in pSNAPf, as a fusion to the N- terminus of the SNAP-tag. Cytochrome C oxidase is located in the inner mitochondrial membrane and is the terminal enzyme of the respiratory chain. The COX8-2-SNAPf fusion protein gives mitochondrial fluorescence when labeled with SNAP-Cell™ substrates. The full sequence and map for pSNAPf-Cox8A can be downloaded.

    The SNAP-tag is a novel tool for protein research, allowing the specific, covalent attachment of virtually any molecule to a protein of interest. The SNAP-tag is a small protein based on human O6-alkylguanine-DNA-alkyltransferase (hAGT). SNAP-tag substrates are derivatives of benzyl purines and benzyl pyrimidines. In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag.

    pSNAPf-Cox8A contains an improved version of SNAP-tag, termed SNAPf. SNAPf displays faster kinetics in in vitro labeling and fast, specific and efficient labeling in live and fixed cell applications, thereby rendering it a desired research tool for analysis of protein dynamics.

    There are two steps to using this system: sub cloning and expression of the protein of interest as a SNAPf fusion, and labeling of the fusion with the SNAP-tag substrate of choice. Expression of the SNAPf-Cox8A fusion protein is described in this document. The labeling of the fusion proteins with SNAP-tag substrates is described in the instructions supplied with SNAP-tag substrates.

    Live HEK293 cells transiently transfected with pSNAPf-Cox8A. Cells were labeled with SNAP-Cell TMR-Star (red) for 15 minutes and counterstained with Hoechst 33342 (blue).


    N9185b_thumb

    Properties and Usage

    Materials Required but not Supplied

    Cell culture media and reagents
    Mammalian cell lines
    Transfection reagents
    SNAP-tag substrates

    Storage Temperature

    -20°C

    Notes

    1. Storage: pSNAPf-Cox8A is supplied in TE buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA) at a concentration of 0.5 µg/µl. Plasmid solutions can be stored at 4°C for up to one week. For long-term storage -20°C is recommended.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.
    1. Cellular Imaging and Analysis FAQs
    1. Expression of SNAPf Fusions (N9185)

    Selection Tools

    Troubleshooting Guides

    Interactive Tools

    Application Notes

    If you generate more plasmid DNA by bacterial transformation, we recommend isolating the plasmid DNA using an endotoxin-free plasmid prep kit prior to transfection into mammalian cells.