DescriptionThis control plasmid contains the gene encoding the Beta-2 adrenergic receptor cloned downstream of the SNAPf coding sequence in pSNAPf as a fusion to the C-terminus of the SNAP-tag. The signal peptide fused to the N-terminus of SNAPf is based on the 5HT3A serotonin receptor. The Beta-2 adrenergic receptor is a member of the G protein coupled receptors and mediates the catecholamine-induced activation of adenylate cyclase through the action of G proteins.
The SNAPf-Beta-2 adrenergic receptor is inserted in the plasma membrane with the SNAPf exposed to the extracellular side of the membrane. When labeled with SNAP-tag substrates, it gives a selective cell membrane fluorescent labeling pattern.
The SNAP-tag is a novel tool for protein research, allowing the specific, covalent attachment of virtually any molecule to a protein of interest. The SNAP-tag is a small protein based on human O6-alkylguanine-DNA-alkyltransferase (hAGT). SNAP-tag substrates are derivatives of benzylguanines and benzylchloropyrimidines. In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag.
There are two steps to using this system: sub cloning and expression of the protein of interest as a SNAPf fusion, and labeling of the fusion with the SNAP-tag substrate of choice. Expression of SNAPf-ADRβ2 fusion protein is described in this document. The labeling of the fusion proteins with SNAP-tag substrates is described in the instructions supplied with SNAP-tag substrates.
DNASU is a central repository for plasmid clones and collections that may also be helpful.
Properties and Usage
Materials Required but not Supplied
- Cell culture media and reagents
- Mammalian cell lines
- Transfection reagents
- SNAP-tag substrates
Legal and Disclaimers
Notice To Buyer/User: The Buyer/User has a non-exclusive license to use this system or any component thereof for RESEARCH AND DEVELOPMENT PURPOSES ONLY. Commercial use of this system or any components thereof requires a license from New England Biolabs, Inc., 240 County Road, Ipswich, MA 01938.
Cellular Imaging and Analysis (i.e., SNAP and CLIP products)
Notice to Buyer/User: The Buyer/User has a non-exclusive license to use this system or any component thereof for RESEARCH AND DEVELOPMENT ONLY. Commercial use of this system or any components thereof requires a license from New England Biolabs, Inc., 240 County Road, Ipswich, MA 01938.
The products and/or their use may be covered by one or more of the following patents and patent applications:
7,939,284 (Methods for Using O6-Alkylguanine-DNA-Alkyltransferases)
7,888,090 (Mutants of O6-Alkylguanine-DNA-Alkyltransferases)
8,163,479 Specific Substrates for O6-Alkylguanine-DNA-Alkyltransferases)
8,178,314 (Pyrimidines Reacting With O6-Alkylguanine-DNA-Alkyltransferases)
PCT/EP2007/057597 (Labeling of Fusion Proteins with Synthetic Probes)
EP07117800 (Drug Delivery)
EP07117802 (Drug Delivery)
EP07120288 (GTPase-Transient Protein Protein Interactions)
These patents and patent applications are owned by Covalys, or owned by the Ecole Polytechnique Fédérale de Lausanne (EPFL) and exclusively licensed to Covalys and NEB.
FuGENE® is a registered trademark of Roche.
Material Safety Datasheets
- Storage: pSNAPf-ADRβ2 is
supplied in TE buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA) at a concentration of
0.5 µg/µl. Plasmid solutions can be stored at 4°C for up to one week. For
long-term storage -20°C is recommended.