The ACP-tag and MCP-tag protein labeling technologies are based on an enzyme-catalysed post-translational modification. Recombinant target protein that is fused to an acyl carrier protein (ACP), is labeled specifically with CoA derivatives, catalyzed by the phosphopantetheinyl transferase ACP Synthase (1). The ACP-tag is small, only 8 kDa (2). MCP-tag, a mutant of ACP-tag, is labeled by the phosphopantetheinyl transferase SFP Synthase but not by ACP Synthase, thereby permitting selective simultaneous labeling in a sample.
Membrane proteins are challenging to study given their hydrophobic nature, generally low native abundance and intrinsic instability (3,4). Regardless, half of all protein drug targets are membrane proteins. For imaging, most fluorescent proteins (i.e. GFP) cannot specifically visualize cell surface subpopulations. However, the ACP-tag/MCP-tag system is a cell surface-specific system since all of its substrates are non-cell permeable. It can specifically label subpopulations of target protein expressed on cell surface using non-cell permeable substrates (5). 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).
- George, N., Pick, H., Vogel, H. et al. (2004) J. Am. Chem. Soc. 126, 8896. PMID: 15264811
- Yin, J., Straight, P.D., McLoughlin, S.M. et al. (2005) Proc. Natl. Acad. Sci. USA 102, 15815. PMID: 16236721
- Lacapère J-J, Pebay-Peyroula E, Neumann J-M, Etchebest C. (2007) Trends Biochem Sci. 32, 259–270. PMID: 17481903
- von Heijne G. (2007) J Intern Med. 261, 543–557. PMID: 17547710
- Keppler, A., Pick, H., Arrivoli, C. et al. (2004) Proc. Natl. Acad. Sci. USA 101, 9955. PMID: 15226507
SNAP-tag® is a registered trademark of New England Biolabs, Inc.
FAQs for ACP/MCP Surface
Protocols for ACP/MCP Surface
- CoA 488 (S9348)
- Instructions for Labeling of Proteins in vitro (S9348)
- Instructions for Use with SFP Synthase (P9302)
- Instructions for Use with ACP Synthase (P9301)
- Labeling of Proteins in vitro (P9301)
- Labeling of Proteins in vitro (P9302)
- Labeling of Proteins in vitro (S9349)
- Labeling of Proteins in vitro (S9350)
- Labeling of Proteins in vitro (S9351)
- Labeling on the Surface of Cells (S9349)
- Labeling on the Surface of Cells (S9350)
- Labeling on the Surface of Cells (S9351)
- 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
Cellular Imaging and 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.
- SNAP-tag® Technologies: Novel Tools to Study Protein Function
- Comparison of SNAP-tag®/CLIP-tag™ Technologies to GFP
- Labeling with SNAP-tag® Technology Troubleshooting Guide
Other Tools & Resources
- 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 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 ACP-tag
SNAP-tag®, CLIP-tag™ and ACP/MCP-tag Substrate Selection Chart
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 firstname.lastname@example.org.
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.