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.
- Cellular Labeling (E9100)
- 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 (S9101)
- Labeling of Proteins in vitro (S9110)
- Labeling of Proteins in vitro (S9103)
- Labeling of Proteins in vitro (S9105)
- Labeling of Proteins in vitro (S9107)
- Labeling of Proteins in vitro (S9109)
- Labeling of Proteins in vitro (S9143)
- Labeling of Proteins in vitro (S9101)
- Labeling Proteins in vitro (S9112)
- Labeling Proteins in vitro (S9124)
- Labeling Proteins in vitro (S9129)
- Labeling Proteins in vitro (S9132)
- Labeling Proteins in vitro (S9134)
- Labeling Proteins in vitro (S9136)
- Use of SNAP-Cell Block with SNAP-Cell Substrates (E9100)
- View the video "Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging" in the Journal of Visualized Experiments (JoVE)
- Cellular Labeling (E9120)
- Labeling Proteins in vitro (E9120)
- Cloning of SNAP-tag Fusions in pSNAPf (N9183)
- Cloning of SNAP-tag Fusions in pSNAP-tag(T7)-2 (N9181)
- Expression of SNAP-tag Fusions (N9181)
- Expression of SNAPf Fusions (N9183)
- Labeling of Proteins in vitro (E9100)
- Use with SNAP-Surface substrates (S9143)
- Cellular Labeling (S9159)
- Labeling of Proteins in vitro (S9159)
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
- Maffei, M., Morelli, C., Graham, E., Patriarca, S., Donzelli, L., Doleschall, B., de Castro, Reis, F., Nocchi, L., Chadick, C.H., Reymond, L., Correa, I.R., Jr., Johnsson, K., Hackett, J.A., Heppenstall, P.A 2019. A ligand based system for receptor specific delivery of proteins Sci Rep. 9(1), PubMedID: 31844114, DOI: 10.1038/s41598-019-55797-1
- 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.
- 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.
- 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.
- 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.
- 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
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at email@example.com.
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.