Cellular Analysis
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  • SNAP Surface

    Membrane proteins are challenging to study given their hydrophobic nature, generally low native abundance and intrinsic instability (1,2).  Regardless, half of all protein drug targets are membrane proteins.  For imaging, most fluorescent proteins (i.e. GFP) cannot specifically visualize cell surface subpopulations.  

    The SNAP-tag® system is based on a DNA repair enzyme, O6-alkylguanine-DNA alkyltransferase (AGT). It allows for multiple substrate options to enable color changes. It is highly temperature and fixation stable and can be used in vitro or in vivo. The substrate consists of two parts: the benzylguanine group and the functional group which can be a fluorophore, biotin or bead. During the labeling reaction the substituted benzyl group covalently attaches to the SNAP-tag releasing guanine. Once the fluorophore is coupled to the desired protein, the label fluorescesces permitting visualization in living or fixed cells.

    SNAP-tag, CLIP-tag™ and cell surface-specific ACP/MCP-tag systems can specifically label subpopulations of target proteins expressed on the cell surface using non-cell permeable substrates (3). This approach permits discrimination of different populations of a cell surface protein: those properly translocated to the plasma membrane from those retained in the secretory pathway or already internalized (e.g. upon ligand binding).

    SNAP-tag® is a registered trademark of New England Biolabs, Inc.
    CLIP-tag™ is a trademark of New England Biolabs, Inc.

    References

    1. Lacapère J-J, Pebay-Peyroula E, Neumann J-M, Etchebest C. (2007) Trends Biochem Sci. 32, 259–270. PMID: 17481903
    2. von Heijne G. (2007) J Intern Med. 261, 543–557. PMID: 17547710
    3. Keppler, A., Pick, H., Arrivoli, C. et al. (2004) Proc. Natl. Acad. Sci. USA, 101, 9955. PMID: 15226507
    • Fluorescent Labeling of COS-7 Expressing SNAP-tag® Fusion Proteins for Live Cell Imaging

      Watch as Chris Provost, of New England Biolabs, performs fluorescent imaging of live COS-7 cells expressing SNAP-tag® fusion proteins.

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    • SNAP-tag® Overview Tutorial

      View an interactive tutorial explaining the mechanism of our SNAP-tag® technologies and reagents available for researchers wishing to study the function and localization of proteins in live or fixed cells.

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    FAQs for SNAP Surface

    Features

    • Clone and express once, then use with a variety of substrates
    • Non-toxic to living cells
    • Wide selection of fluorescent substrates
    • Highly specific covalent labeling
    • Simultaneous dual labeling

    Applications

    • Simultaneous dual protein labeling on the surface of live cells
    • Protein localization and translocation
    • Pulse-chase experiments
    • Receptor internalization studies
    • Selective cell surface labeling
    • Protein pull-down assays
    • Protein detection in SDS-PAGE
    • Flow cytometry
    • High throughput binding assays in microtiter plates
    • Biosensor interaction experiments
    • FRET-based binding assays
    • Single molecule labeling
    • Super-resolution microscopy

    Protein Labeling with SNAP-tag- and CLIP-tag

    The SNAP- (gold) or CLIP-tag (purple) is fused to the protein of interest (blue). Labeling occurs through covalent attachment to the tag, releasing either a guanine or a cytosine moiety.

    SNAP-tag®, CLIP-tag™ and ACP/MCP-tag Substrate Selection Chart

    NEB offers a large selection of fluorescent labels (substrates) for SNAP-, CLIP-, ACP- and MCP-tag fusion proteins.