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How it works:
ACP-tag is fused to the extracellular part of the membrane protein, and the fusion protein is expressed. ACP-Synthase and ACP substrate are used for selective fluorescent labeling of tagged protein exposed on the cell membrane.
Advantages:
- Clear signal as the substrates are not cell-permeable and only surface exposed tag is accessible to the labling synthase.
- Fluorescent label can be selected to best fit instrumentation.
- Recently the ACP-tag was used successfully to investigate dimerization of the NK1 receptor by a FRET-based assay. It was demonstrated that NK1-receptors are monomeric and reside in membrane microdomains of live cells.
Note: SNAP-tag can also be used for selective cell-surface labeling, see Receptor Internalization
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| CHO-K1 cells expressing ACPwt-tag-NK-1R fusion, labeled with CoA-488 (green), counterstained with Hoechst 33342 (blue). | | COS7 cells expressing ACPwt-tag-GPI anchor fusion, labeled with CoA-547 (red), counterstained with Hoechst 33342 (blue). |
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