pACP-ADRβ2 Control Plasmid

  • Catalog # N9321 was discontinued on January 01, 2013
  • Product Information
    This control plasmid contains the gene encoding the Beta-2 Adrenergic Receptor cloned as a fusion to the C-terminus of the ACP-tag. The signal peptide fused to the N-terminus of ACP-tag 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 ACP-Beta-2 Adrenergic Receptor is inserted in the plasma membrane with the ACP-tag exposed to the extracellular side of the membrane. When labeled with CoA substrates, it gives a selective cell membrane fluorescence labeling pattern.The full sequence and map for pACP-ADRβ2 can be downloaded here.

    The ACP-tag is a small protein tag (8 kDa) based on the acyl carrier protein (ACP). It allows the specific, covalent attachment of virtually any molecule to a protein of interest. ACP-tag substrates are derivatives of coenzyme A (CoA). In the labeling reaction, the substituted phosphopantetheine group of CoA is covalently attached to a conserved serine residue of the ACP-tag by a phosphopantetheine transferase (ACP or SFP Synthase). Having no cysteines, the ACP-tag is particularly suited for specifically labeling cell-surface proteins, and should be useful for labeling secreted proteins with disulfide bridges such as antibodies.

    There are two steps to using this system: subcloning and expression of the protein of interest as an ACP-tag fusion, and labeling of the fusion protein with the CoA substrate of choice. The cloning and expression of ACP-tag protein fusions is described in documents provided with the pACP-tag(m)-2 cloning plasmid. The labeling of fusion proteins with CoA substrates is described in the documentation supplied with CoA substrates and ACP or SFP Synthase.

    Live U-2 0S cells transiently transfected with pACP-ADRβ2. Cells were labeled with CoA 547 using SFP Synthase for 30 minutes and imaged by confocal microscopy.
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