UNIT 26.5 TimeSTAMP Tagging of Newly Synthesized Proteins
Published Online: 1 FEB 2010
Copyright © 2010 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Protein Science
How to Cite
Lin, M. Z. and Tsien, R. Y. 2010. TimeSTAMP Tagging of Newly Synthesized Proteins. Current Protocols in Protein Science. 59:26.5:26.5.1–26.5.11.
- Published Online: 1 FEB 2010
- Published Print: FEB 2010
The ability to quantify or visualize newly synthesized proteins has important uses in cell biology. For example, a researcher may wish to quantify basal or inducible rates of translation of a specific gene of interest, or detect subcellular locations of newly synthesized copies of a protein in order to study the role of new protein synthesis in the growth of specialized cellular structures. In this unit, the TimeSTAMP method for labeling of newly synthesized copies of a protein of interest is described. In the TimeSTAMP method, the experimenter expresses a protein of interest as a fusion with a cis-acting protease and an epitope tag, both of which are removed by default protease activity. Addition of a specific protease inhibitor then allows preservation of the tag on subsequently synthesized proteins. Finally, the tag is detected by immunological methods. Curr. Protoc. Protein Sci. 59:26.5.1-26.5.11. © 2010 by John Wiley & Sons, Inc.
- protein synthesis;
- conditional labeling