Product Class: Vector

pSNAPf -ADRβ2 Control Plasmid

Catalog #SizeConcentration
N9184S20 μg0.5 mg/ml

Description

This 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. 

pSNAPf-ADRβ2 contains an improved version of SNAP-tag, termed SNAPf. SNAPf displays faster kinetics in in vitro labeling and fast, specific and efficient labeling in live and fixed cell applications, thereby rendering it a desired research tool for analysis of protein dynamics.

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.

Live HEK293 cells transiently transfected with pSNAPf-ADRβ2. Cells were labeled with SNAP-Surface™ 488 (green) for 15 minutes.

DNASU is a central repository for plasmid clones and collections that may also be helpful.



N9184b

Properties and Usage

Materials Required but not Supplied

  • Cell culture media and reagents 
  • Mammalian cell lines 
  • Transfection reagents 
  • SNAP-tag substrates

Storage Temperature

-20°C

Supporting Documents

Material Safety Datasheets

The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Notes

  1. 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.
  1. Cellular Imaging and Analysis FAQs

Application Notes

If you generate more plasmid DNA by bacterial transformation, we recommend isolating the plasmid DNA using an endotoxin-free plasmid prep kit prior to transfection into mammalian cells.