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Related Concept Videos

Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

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Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
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DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
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Protein Dynamics in Living Cells01:19

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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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Related Experiment Video

Updated: May 3, 2026

Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging
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Site-specific protein labeling with SNAP-tags.

Nelson B Cole1

  • 1Laboratory of Cell Biology, National Heart, Lung, and Blood Institute/National Institutes of Health (NHLBI/NIH), Bethesda, Maryland.

Current Protocols in Protein Science
|February 11, 2014
PubMed
Summary

Site-specific protein labeling using SNAP-tag technology enables precise study of protein function in living cells. This method utilizes O(6)-benzylguanine derivatives for versatile labeling with various reporters, including releasable probes.

Keywords:
SNAP-tagcell biologychemical labelingendocytosisimaging

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Site-specific protein labeling is crucial for understanding protein function in vivo.
  • Small peptide and protein tags offer advantages over fluorescent proteins for chemical labeling.
  • SNAP-tag, derived from human O(6)-alkylguanine-DNA-alkyltransferase, facilitates covalent labeling with O(6)-benzylguanine (BG) derivatives.

Purpose of the Study:

  • To describe basic strategies for labeling SNAP-tag fusion proteins.
  • To enable live cell imaging and in vitro analysis of protein function.
  • To introduce a releasable SNAP-tag probe for controlled reporter cleavage.

Main Methods:

  • Utilizing the SNAP-tag's self-labeling reaction with modified O(6)-benzylguanine (BG) probes.
  • Employing a variety of fluorophores and reporter compounds attached to BG derivatives.
  • Implementing a releasable probe strategy for chemical cleavage of the label.

Main Results:

  • Demonstrated efficient and specific labeling of SNAP-tag fusion proteins.
  • Showcased versatility of BG derivatives for diverse reporter conjugation.
  • Successfully developed and described a releasable SNAP-tag probe system.

Conclusions:

  • SNAP-tag technology provides a flexible and efficient platform for site-specific protein labeling.
  • The described methods support both live-cell imaging and in vitro protein analysis.
  • Releasable probes offer advanced control over reporter presence in biological systems.