pACP-tag(m)-2 Vector is a mammalian expression plasmid encoding ACPwt, an ACP-tag protein, which is expressed under control of the CMV promoter. The expression plasmid has an IRES (Internal Ribosome Entry Site) and a neomycin resistance gene downstream of the ACP-tag for the efficient selection of stable transfectants. This plasmid is intended for the cloning and transient or stable expression of ACP-tag protein fusions in mammalian cells. pACP-tag(m)-2 contains two multiple cloning sites to allow cloning of the fusion partner as a fusion to the C-terminus of the ACP-tag and an appropriate signal peptide to the N-terminus of the ACP-tag.
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. In this document, the cloning and expression of ACP-tag protein fusions is described. The labeling of fusion proteins with CoA substrates is described in the documentation supplied with CoA substrates and ACP or SFP Synthase.
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