Localization and Affinity Purification (LAP) Tagging

To facilitate cell biological analysis and proteomics in higher eukaryotes, we have developed the LAP tag which combines the advantages of generating a fluorescent protein fusion such as GFP or mRFP with flexibility of the tandem affinity purification (TAP) tag. The N-terminal and C-terminal LAP tags and their corresponding purification strategies are diagrammed below. For complete plasmid sequences and additional information, click here.

In human cells, the LAP tag faciliates the generation of a stable cell line which can be sorted by FACS to obtain clonal cell lines expressing moderate amounts of the fusion protein. This cell line can be used for a variety of cell biological analyses. This cell line can also be used for an efficient affinity purification to identify interacting proteins. An example is shown below: