SNAP- and CLIP-tag protein labeling systems enable the specific, covalent attachment of virtually any molecule to a protein of interest. There are two steps to using this system: cloning and expression of the protein of interest as a SNAP-tag® fusion, and labeling of the fusion with the SNAP-tag substrate of choice. The SNAP-tag is a small protein based on human O6-alkylguanine-DNA-alkyltransferase (hAGT), a DNA repair protein. SNAP-tag substrates are dyes, fluorophores, biotin, or beads conjugated to guanine or chloropyrimidine leaving groups via a benzyl linker. In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag. CLIP-tag™ is a modified version of SNAP-tag, engineered to react with benzylcytosine rather than benzylguanine derivatives. When used in conjunction with SNAP-tag, CLIP-tag enables the orthogonal and complementary labeling of two proteins simultaneously in the same cells.
SNAP-tag® is a registered trademark of New England Biolabs, Inc.
CLIP-tag™ is a trademark of New England Biolabs, Inc.
SNAP-tag® Technologies: Tools to Study Protein Function
Read about the NEB’s set of protein tools for the specific labeling (SNAP-, CLIP-, ACP- and MCP-tags) of fusion proteins.
- Cellular Imaging & Analysis Brochure
- Comparison of SNAP-tag®/CLIP-tag™ Technologies to GFP
- Labeling with SNAP-tag® Technology Troubleshooting Guide
- Genome-wide profiling of nuclease protected domains reveals physical properties of chromatin
- In Vitro Reconstitution of Thermococcus Species 9°N Okazaki Fragment Maturation (2015)
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- Erhardt, S. et al. 2008. Genome-wide analysis reveals a cell cycle-dependent mechanism controling centromere propagation J. Cell Biol.. 183 , PubMedID: 19047461, DOI:
- Howland S.W. et al. 2008. Inducing efficient cross-priming using antigen-coated yeast particles J. Immunother.. 31 , PubMedID: 18600183, DOI:
- Southwell, A.L. et al. 2008. Intrabodies binding the proline-rich domains of mutant huntingtin increase its turnover and reduce neurotoxicity J. Neurosci. . 28, PubMedID: 18768695, DOI:
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- Degorce F. et al. 2009. HTRF: A technology tailored for drug discovery - a review of theoretical aspects and recent applications Curr. Chem. Genomics . 3 , PubMedID: 20161833, DOI:
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- Kindermann M. et al. 2004. Synthesis and characterization of bifunctional probes for the specific labeling of fusion proteins Bioorg. Med. Chem. Lett. . 14, PubMedID: , DOI:
- Hoskins, A. et al. 2011. Ordered and dynamic assembly of single spliceoseoms Science . 331 , PubMedID: 21393538, DOI:
- Eckhardt, M. et al. 2011. A SNAP-tagged detivative of HIV-1 - A versatile tool to study virus-cell interactions PLoS One . , PubMedID: 21799764, DOI: 10.137/journal. P One .0022007
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- Keppler A. et al. 2006. Fluorophores for live cell imaging of AGT fusion proteins across the visible spectrum BioTechniques . 41, PubMedID: 16925018, DOI:
- Krayl M. et al. 2006. Fluorescence-mediated analysis of mitochondrial preprotein import in vitro Anal. Biochem. . 335, PubMedID: 16750157, DOI:
- Tirat A. et al. 2006. Evaluation of two novel tag-based labeling technologies for site-specific modification of proteins Int. J. Biol. Macromol.. 39, PubMedID: 16503347, DOI:
- Heinis C. et al. 2006. Evolving the substrate specificity of O6 alkylguanine DNA alkyltransferase through loop insertion for applications in molecular imaging ACS Chem Biol. . 1, PubMedID: 17168553, DOI:
- Gronemeyer T. et al. 2006. Adding value to fusion proteins through covalent labeling Curr. Opin. Biotechn. . 16 , PubMedID: 15967656, DOI:
- Johnsson N. et al. 2005. Protein chemistry on the surface of living cells Chembiochem. . 6 , PubMedID: 15558647, DOI:
- Regoes A. et al. 2005. SNAP-tag mediated live cell labeling as an alternative to GFP in anaerobic organisms BioTechniques . 39, PubMedID: , DOI:
- Juillerat A. et al. 2005. Engineering substrate specificity of O6-alkylguanine-DNA alkyltransferase for specific protein labeling in living cells ChemBioChem . 6, PubMedID: 15934048, DOI:
- Srikun, D. et al. 2010. Organelle-targetable fluorescent probes for imaging hydrogen peroxide in living cells via SNAP-tag protein labeling J. Am. Chem. Soc. . 132 , PubMedID: 20201528, DOI:
- Maurel D. et al. 2010. Photoactivatable and photoconvertible fluorescent probes for protein labeling ACS Chem. Biol. Asap . , PubMedID: 20218675, DOI:
- Alvarez-Curto J. et al. 2010. Ligand regulation of the quaternary organization of cell surface M3 muscarinic acetylcholine receptors analyzed by fluorescence resonance energy transfer (FRET) imaging and homogenous time-resolved FRET J. Biol. Chem. . 285 , PubMedID: 20489201, DOI:
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- Campos, C. et al. 2010. Labeling cell structures and tracking cell lineage in zebrafish using SNAP-Tag Dev. Dynamics . 240 , PubMedID: 21360787, DOI:
- Kamiya M. and Johnsson K. 2010. Localizable and Highly Sensitive Calcium Indicator Based on a BODIPY Fluorophore Anal. Chem. . 82 , PubMedID: 20590099, DOI:
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- Hein B. et al. 2010. Stimulated emission depletion nanoscopy of living cells using SNAP-Tag fusion proteins Biophys. J. . 98 , PubMedID: 20074516, DOI:
- Kampmeier, F. et al. 2010. Rapid optical imaging of EGF receptor expression with a single-chain antibody SNAP-tag fusion protein Eur. J. Med. Mol. Imaging . , PubMedID: 20449589, DOI: 10.007/S00259-010-1482-5
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- Liu E and Bruner S. D. 2007. Rational manipulation of carrier-domain geometry in nonribosomal peptide synthetases ChemBioChem. . 8, PubMedID: 17335097, DOI:
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- Generosi J. et al. 2008. Photobleaching-free infrared near-field microscopy localizes molecules in neurons J. App. Phys. . 104, PubMedID: , DOI:
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- Schulz C. and Köhn M. 2008. Simultaneous protein tagging in two colors Chemistry & Biology . 15, PubMedID: 18291310, DOI:
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- Generosi J. et al. 2008. AMPA receptor imaging by infrared scanning near-field optical microscopy Physica Status Solidi C: Current Topics in Solid State Physics . 5, PubMedID: , DOI:
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- Neugart F. et al. 2009. Detection of ligand-induced CNTF receptor dimers in living cells by fluorescence cross correlation spectroscopy Biochim. Biophys. Acta. . 1788 , PubMedID: 19482006, DOI:
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- Kindermann M. et al. 2003. Covalent and selective immobilization of fusion proteins JACS . 125, PubMedID: 12822993, DOI:
- La Clair, J.J. et al. 2004. Manipulation of carrier proteins in antibiotic biosynthesis Chem. Biol. . 11, PubMedID: 15123281, DOI:
- George N. et al. 2004. Specific labeling of cell surface proteins with chemically diverse compounds J .Am. Chem. Soc. . 126, PubMedID: 15264811, DOI:
- Huber W. et al. 2004. SPR-based interaction studies with small molecular weight ligands using hAGT fusion proteins Anal. Biochem. . 333, PubMedID: 15450803, DOI:
- Sielaff I. et al. 2006. Protein function microarrays based on self-immobilizing and self-labeling fusion proteins ChemBioChem.. 7, PubMedID: 16342318, DOI:
- Prummer M. et al. 2006. Post-translational covalent labeling reveals heterogeneous mobility of individual G protein-coupled receptors in living cells ChemBioChem . 7, PubMedID: 16607667, DOI:
- Jacquier V. et al. 2006. Visualizing receptor trafficking in living PNAS . 103, PubMedID: 16980412, DOI:
- Jongsma M.A., Litjens R. H. 2006. Self-assembling protein arrays on DNA chips by auto-labeling fusion proteins with a single DNA address Proteomics . 6, PubMedID: 16596705, DOI:
- Meyer B.H. et al. 2006. Covalent labeling of cell-surface proteins for in vivo FRET studies FEBS Letters . 580, PubMedID: 16497304, DOI:
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- Cravatt B.F. 2005. Live chemical reports from the cell surface Chem. Biol. . 12, PubMedID: 16183017, DOI:
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- Yin J. et al. 2005. Single-cell FRET imaging of transferrin receptor trafficking dynamics by Sfp-catalyzed, site-specific protein labeling Chem. Biol . 12, PubMedID: 16183024, DOI:
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- Mosiewicz, K. A. et al. 2010. Phosphopantetheinyl Transferase-Catalyzed Formation of Bioactive Hydrogels for Tissue Engineering J. Am. Chem. Soc. . 132, PubMedID: 20373804, DOI:
- Engin S. et al. 2010. Benzylguanine Thiol self-assembled monolayers for the immobilization of SNAP-tag proteins on microcontact-printed surface structures Langmuir . ASAP, PubMedID: 20369837, DOI:
- Waichman S. et al. 2010. Functional Immobilization and Patterning of Proteins by an Enzymatic Transfer Reaction Anal. Chem. . 82 , PubMedID: 20092261, DOI:
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- 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
Lukinavičius, G. et al. (2015) "Fluorescent labeling of SNAP-tagged proteins in cells" Methods Mol. Biol. 1266, 107-118.
Corrêa Jr., I. R. (2015) "Considerations and protocols for the synthesis of custom protein labeling probes" Methods Mol. Biol. 1266, 55-79.
Corrêa Jr., I. R. (2014) "Live-cell reporters for fluorescence imaging" Curr. Opin. Chem. Biol. 20, 36-45.
Bosch, P. J. et al. (2014) "Evaluation of fluorophores to label SNAP-tag fused proteins for multicolor single-molecule tracking microscopy in live cells" Biophys. J. 107, 803-814.
Smith, B. A. et al. (2013) "Three-color single molecule imaging shows WASP detachment from Arp2/3 complex triggers actin filament branch formation" eLife 2, e01008.
Jaiswal, R. et al. (2013) "The Formin Daam1 and Fascin Directly Collaborate to Promote Filopodia Formation" Curr. Biol. 23, 1373-1379.
Breitsprecher, D. et al. (2012) "Rocket Launcher Mechanism of Collaborative Actin Assembly Defined by Single-Molecule Imaging" Science 336, 1164-1168.
Hoskins, A. A. et al. (2011) "Ordered and dynamic assembly of single spliceosomes." Science 331 (6022), 1289-1295.
Zhao, Z. W. et al. (2014) "Spatial organization of RNA polymerase II inside a mammalian cell nucleus revealed by reflected light-sheet superresolution microscopy" Proc. Natl. Acad. Sci. USA 111, 681-686.
Lukinavičius, G. et al. (2013) "A near-infrared fluorophore for live-cell super-resolution microscopy of cellular proteins" Nat. Chem. 5, 132-139.
Jones, S. A. et al. (2011) "Fast, three-dimensional super-resolution imaging of live cells." Nat. Methods 8, 499-505.
Klein, T. et al. (2011) "Live-cell dSTORM with SNAP-tag fusion proteins." Nat. Methods 8, 7-9.
Pellett, P. A. et al. (2011) "Two-color STED microscopy in living cells." Biomed. Opt. Expr. 2, 2364-2371
Hein, B. et al. (2010) "Stimulated Emission Depletion Nanoscopy of Living Cells Using SNAP-Tag Fusion Proteins." Biophys. J. 98, 158-163.
Tissue and Animal Imaging:
Yang, G. et al. (2015) "Genetic targeting of chemical indicators in vivo" Nat. Methods 12, 137-139.
Kohl, J. et al. (2014) "Ultrafast tissue staining with chemical tags" Proc. Natl. Acad. Sci. USA 111, E3805-E3814.
Ivanova, A. et al. (2013) "Age-dependent labeling and imaging of insulin secretory granules" Diabetes 62, 3687-3696.
Gong, H. et al. (2012) "Near-Infrared Fluorescence Imaging of Mammalian Cells and Xenograft Tumors with SNAP-Tag" PLoS ONE 7(3): e34003.
Bojkowska K. et al. (2011) "Measuring in vivo protein half-life." Chem. Biol. 18, 805-815.
Cell-Surface Protein Labeling and Internalization Analysis:
Bitsikas, V. et al. (2014) "Clathrin-independent pathways do not contribute significantly to endocytic flux" eLife 3, e03970.
Jaensch, N. et al. (2014) "Stable Cell Surface Expression of GPI-Anchored Proteins, but not Intracellular Transport, Depends on their Fatty Acid Structure" Traffic 15, 1305-1329.
Cole, N. B. and Donaldson, J. G. (2012) "Releasable SNAP-tag Probes for Studying Endocytosis and Recycling" ACS Chem. Biol. 7, 464-469.
Rošić, S. et al. (2014) "Repetitive centromeric satellite RNA is essential for kinetochore formation and cell division" J. Cell Biol. 207, 335-349.
Stoops, E. H. et al. (2014) "SNAP-Tag to Monitor Trafficking of Membrane Proteins in Polarized Epithelial Cells" Methods Mol. Biol. 1174, 171-182.
Bordor, D. L. et al. (2012) "Analysis of Protein Turnover by Quantitative SNAP-Based Pulse-Chase Imaging" Curr. Protoc. Cell Biol. 55, 8.8.1-8.8.34.
Register, A. C. et al. (2014) "SH2-Catalytic Domain Linker Heterogeneity Influences Allosteric Coupling across the SFK Family" Biochemistry 53, 6910-6923.
Shi, G. et al. (2012) "SNAP-tag based proteomics approach for the study of the retrograde route" Traffic 13, 914-925.
Bieling, P. et al. (2010) "A minimal midzone protein module controls formation and length of antiparallel microtubule overlaps" Cell 142, 420-432.
Protein-Protein and Protein-Ligand Interactions:
Griss, R. et al. (2014) "Bioluminescent sensor proteins for point-of-care therapeutic drug monitoring" Nat. Chem. Biol. 10, 598-603.
Chidley, C. et al. (2011) "A yeast-based screen reveals that sulfasalazine inhibits tetrahydrobiopterin biosynthesis." Nat. Chem. Biol. 7, 375-383.
Gautier A. et al. (2009) "Selective Cross-Linking of Interacting Proteins using Self-Labeling Tags" J. Am. Chem. Soc. 131, 17954-17962.
Maurel D. et al. (2008) "Cell-surface protein-protein interaction analysis with time-resolved FRET and SNAP-tag technologies: application to GPCR oligomerization." Nat. Methods 5, 561-567.
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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.
Watch as Chris Provost, of New England Biolabs, performs fluorescent imaging of live COS-7 cells expressing SNAP-tag® fusion proteins.
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.