Cellular Analysis
  • My NEB
  • Print
  • PDF
  • Cell Imaging

    Cell imaging analysis can use fluorescent dyes, fluorophore labeled molecules or recombinant protein plasmid systems. Recombinant protein labeling systems and  bioluminescent reporter systems are among the most sensitive fluorescence methods for imaging expression, transport, co-localization and degradation in either fixed or living cells. Protein labeling systems offer many advantages. For example, color changes can be easily implemented by using different substrates. Protein labeling systems can involve the use of tag-specific antibodies or antibodies to separate epitopes engineered into a plasmid tag system for detection. Protein labeling systems can be used with non-cell permeable substrates to enable the specific imaging of cell surface targets. This strategy is not possible with bioluminescent recombinant systems.  In living cells, protein labeling substrates can be introduced and followed in cells over time. Two separate cellular targets can also be imaged simultaneously, using protein labeling systems with mutually exclusive, tag specific fluorescent substrates. Cellular functions and structures can be visually detected using methods such as wide-field fluorescence, confocal, time-resolved and fluorescence resonance energy transfer (FRET) microscopy, or cell based high content assays.  Given cellular target characteristics and the array of possible detection methods, choosing an optimal fluorescence system for cellular imaging is key to experimental design.  

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

    Featured Products

    Cell Imaging includes these areas of focus:

    SNAP Companion
    SNAP Cell
    SNAP Surface
    CLIP Cell
    CLIP Surface
    CLIP Companion
    ACP/MCP Surface
    ACP/MCP Companion

    FAQs for Cell Imaging

    Protocols for Cell Imaging

    Features of SNAP-tag/CLIP-tag

    • 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 of SNAP-tag/CLIP-tag

    • Simultaneous dual protein labeling inside 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

    Advantages of ACP/MCP-tag

    • Small - Expressed tag is only 8 kDa (77aa)
    • Versatile - ACP- and MCP-tagged fusions can be co-expressed and sequentially labeled for two color applications on cell surfaces
    • Specific - Components remain exclusively extracellular, preventing intracellular labeling
    • Precise - Label is covalently bound under biological conditions in a defined position
    • Non-toxic - Substrates are non-toxic to living cells
    • Selection - Choice of substrates available, including 488, 547, 647 nm and biotin

    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.

    Protein Labeling with ACP-tag

    ACP-tag (red) fused to the protein of interest (blue) is labeled in the presence of a required synthase.

    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.