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.
FAQs for SNAP Surface
Protocols for SNAP Surface
- Cellular Labeling (E9100)
- Cellular Labeling (E9120)
- Cellular Labeling (S9101)
- Cellular Labeling (S9103)
- Cellular Labeling (S9105)
- Cellular Labeling (S9107)
- Cellular Labeling (S9109)
- Cellular Labeling (S9110)
- Cellular Labeling (S9112)
- Cellular Labeling (S9124)
- Cellular Labeling (S9129)
- Cellular Labeling (S9132)
- Cellular Labeling (S9134)
- Cellular Labeling (S9136)
- Cellular Labeling (S9159)
- Cloning of SNAP-tag Fusions in pSNAPf (N9183)
- Cloning of SNAP-tag Fusions in pSNAP-tag(T7)-2 (N9181)
- Expression of SNAPf Fusions (N9183)
- Expression of SNAP-tag Fusions (N9181)
- Labeling of Proteins in vitro (S9107)
- Labeling of Proteins in vitro (S9110)
- Labeling of Proteins in vitro (S9101)
- Labeling of Proteins in vitro (S9103)
- Labeling of Proteins in vitro (S9105)
- Labeling of Proteins in vitro (S9109)
- Labeling of Proteins in vitro (S9143)
- Labeling of Proteins in vitro (E9100)
- Labeling of Proteins in vitro (S9159)
- Labeling Proteins in vitro (S9132)
- Labeling Proteins in vitro (S9112)
- Labeling Proteins in vitro (S9129)
- Labeling Proteins in vitro (S9134)
- Labeling Proteins in vitro (S9136)
- Labeling Proteins in vitro (E9120)
- Labeling Proteins in vitro (S9124)
- Use of SNAP-Cell Block with SNAP-Cell Substrates (E9100)
- Use with SNAP-Surface substrates (S9143)
- View the video "Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging" in the Journal of Visualized Experiments (JoVE)
Application Notes SNAP Surface
- Labeling and Imaging of Cell Surface Receptors Mediated by SNAP-tag®
- Labeling of Escherichia coli Expressed SNAP-tag® Fusion Proteins
- Simultaneous Fluorescent Labeling of Proteins in Living Cells
- Simultaneous Labeling of Two Proteins in Live Cells
- SNAPf based pulse labeling for analysis of protein turnover in living cells
SNAP-tag® Technologies: Novel Tools to Study Protein Function
Cellular Imaging & Analysis Brochure
The Cellular Imaging and Analysis brochure provides information on the labeling technologies offered by NEB for studying the function and localization of proteins in cells.
- Comparison of SNAP-tag®/CLIP-tag™ Technologies to GFP
- Labeling with SNAP-tag® Technology Troubleshooting Guide
Other Tools & Resources
- Elena Shvets, Vassilis Bitsikas, Gillian Howard, Carsten Gram Hansen, Benjamin J Nichols 2015. Dynamic caveolae exclude bulk membrane proteins and are required for sorting of excess glycosphingolipids Nat Commun. 6, PubMedID: 25897946, DOI: 10.1038/ncomms7867.
- Margaret L Rodgers, Joshua Paulson, Aaron A Hoskins 2015. Rapid isolation and single-molecule analysis of ribonucleoproteins from cell lysate by SNAP-SiMPull RNA. 21(5), PubMedID: 25805862, DOI: 10.1261/rna.047845.114
- Meron Mengistu, Krishanu Ray, George K Lewis, Anthony L DeVico 2015. Antigenic properties of the human immunodeficiency virus envelope glycoprotein gp120 on virions bound to target cells PLoS Pathog. 11(3), PubMedID: 25807494, DOI: 10.1371/journal.ppat.1004772.
- Gabriele Fuchs, Alexey N Petrov, Caleb D Marceau, Lauren M Popov, Jin Chen, Sen E O'Leary, Richard Wang, Jan E Carette, Peter Sarnow, Joseph D Puglisi 2015. Kinetic pathway of 40S ribosomal subunit recruitment to hepatitis C virus internal ribosome entry site Proc Natl Acad Sci U S A. 112(2), PubMedID: 25516984, DOI: 10.1073/pnas.1421328111.
- Juri Nio Bach, Marc Bramkamp 2015. Dissecting the molecular properties of prokaryotic flotillins PLoS One. 10(1), PubMedID: 25635948, DOI: 10.1371/journal.pone.0116750.
Publications related to SNAP Surface
- 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
SNAP-tag®, CLIP-tag™ and ACP/MCP-tag Substrate Selection Chart
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.