Protein Labeling
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.
CLIP-tag™ is a trademark of New England Biolabs, Inc.
SNAP-tag® is a registered trademark of New England Biolabs, Inc.
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FAQs for Protein Labeling
Protocols for Protein Labeling
- Fusion Constructs (E6901)
- Labeling of Proteins in vitro (S9159)
- Cellular Labeling (S9159)
- Cellular Labeling (S9102)
- Labeling of Proteins in vitro (S9102)
- Cellular Labeling (S9233)
- Labeling Proteins in vitro (S9129)
- Labeling of Proteins in vitro (S9103)
- Instructions for Use with SFP Synthase (P9302)
- Primer Design for Restriction Enzyme Cloning (E6901)
- Preparation of Media and Solutions (E6901)
- Labeling Proteins in vitro (S9132)
- Labeling Proteins in vitro (S9136)
- Labeling Proteins in vitro (S9124)
- Use of CLIP-Cell Block with CLIP-Cell Substrates (E9230)
- Use of SNAP-Cell Block with SNAP-Cell Substrates (E9100)
- Simplified Expression and Purification Protocol (E6901)
- Cellular Labeling (S9219)
- Cellular Labeling (S9232)
- Cellular Labeling (S9217)
- Cellular Labeling (S9234)
- Cellular Labeling (S9107)
- Cellular Labeling (S9112)
- Cellular Labeling (S9124)
- Cellular Labeling (S9104)
- Cellular Labeling (S9105)
- Cellular Labeling (S9109)
- Cellular Labeling (S9134)
- Cellular Labeling (S9136)
- Cellular Labeling (S9129)
- Cellular Labeling (S9132)
- Construction of the Fusion Plasmid (E6901)
- CoA 488 (S9348)
- Cellular Labeling (S9103)
- Cellular Labeling (E9100)
- Cellular Labeling (E9230)
- Affinity Purification and On-column Cleavage (E6901)
- Labeling of Proteins in vitro (S9107)
- Labeling of Proteins in vitro (S9217)
- Labeling of Proteins in vitro (P9302)
- Labeling of Proteins in vitro (S9105)
- Instructions for Use with ACP Synthase (P9301)
- Cellular Labeling (S9101)
- Labeling on the Surface of Cells (S9349)
- Labeling of Proteins in Solution (E9230)
- Labeling of Proteins in vitro (S9101)
- Labeling Proteins in vitro (S9134)
- Labeling on the Surface of Cells (S9350)
- Labeling Proteins in vitro (S9112)
- Labeling of Proteins in vitro (S9219)
- Labeling of Proteins in vitro (S9234)
- Labeling of Proteins in vitro (S9104)
- Instructions for Labeling of Proteins in vitro (S9348)
- Labeling of Proteins in vitro (S9233)
- Labeling of Proteins in vitro (S9109)
- Labeling of Proteins in vitro (S9232)
- Labeling of Proteins in vitro (P9301)
- Fusion Protein Expression (E6901)
- Labeling Proteins in vitro (E9200)
- Labeling Proteins in vitro (E9120)
- Instructions for Cellular Labeling (E9200)
- Labeling of Proteins in vitro (E9100)
- Labeling of Proteins in vitro (S9349)
- Labeling of Proteins in vitro (S9350)
- Cellular Labeling (E9120)
- Use with SNAP-Cell Substrates (S9106)
- Protocol for Anti-SNAP-tag® Antibody (Polyclonal) (P9310)
- Labeling Proteins in vitro (S9147)
- Labeling SNAP-tag Fusion Proteins with BG-substrates (S9151)
- Labeling SNAP-tag Purified Protein In Vitro (P9312)
- Labeling SNAP-tag Fusion Proteins with BG-substrates (S9150)
- Use with CLIP-tag substrates (S9220)
- Cellular Labeling (S9110)
- Cloning of CLIP-tag Fusions in pCLIPf (N9215)
- Labeling Mammalian Cell Lysates (S9147)
- Labeling CLIP-tag Fusion Proteins with BC-substrates (S9236)
- Labeling of Proteins in vitro (S9110)
- Labeling of Proteins in vitro (S9106)
- Labeling of Proteins in vitro (S9351)
- Labeling on the Surface of Cells (S9351)
- Labeling of Proteins in vitro (S9220)
- Labeling of Proteins in vitro (S9143)
- Labeling of Proteins in vitro (S9221)
- Expression of CLIP-tag Fusions (N9215)
- Use SNAP-Capture Pull Down Resin (S9144)
- Reaction Conditions for Chemical Coupling (S9153)
- Reaction Conditions for Chemical Coupling (S9148)
- Cellular Labeling (S9221)
- Use with SNAP-Surface substrates (S9143)
- Protocol for Labeling SNAP-tag Fusion Proteins with BG-substrates (S9153)
- Labeling SNAP-tag Fusion Proteins with BG-substrates (S9148)
- Expression of SNAPf Fusions (N9183)
- 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)
- Reaction Conditions for Chemical Coupling (S9151)
- Reaction Conditions for Chemical Coupling (S9236)
- Reaction Conditions for Chemical coupling (S9150)
- View the video "Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging" in the Journal of Visualized Experiments (JoVE)
- Use SNAP-Capture Magnetic Beads (S9145)
Application Notes for Protein Labeling
- Intein-Mediated Protein Ligation (IPL) and Labeling with the IMPACT™ Kit
- 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
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SNAP-tag® Technologies: Novel Tools to Study Protein Function
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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
Feature Articles
Brochures
Selection Tools
Troubleshooting Guides
Features
- 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
- 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
Protein Labeling with SNAP-tag and CLIP-tag
Protein Labeling with ACP-tag
Videos
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Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging
Watch as Chris Provost, of New England Biolabs, performs fluorescent imaging of live COS-7 cells expressing SNAP-tag® fusion proteins.
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SNAP-tag Overview Tutorial
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.
