AI Article Synopsis
- Epitope tagging is a method used to study cellular components by revealing their location, interactions, and functions.
- The process of tag-based affinity purification produces a sample containing the target molecule along with associated and background proteins and nucleic acids, influenced by the specific protocol used.
- The chapter outlines how to design an affinity purification experiment and discusses common techniques for detecting proteins, RNA, and DNA in Tetrahymena thermophila.
Article Abstract
Epitope tagging is a powerful approach used to enable investigations of a cellular component by elucidating its localization, interaction partners, and/or activity targets. Successful tag-based affinity purification yields a mixture of the molecule of interest, associated proteins and nucleic acids, and nonspecific background proteins and nucleic acids, many of which can depend on details of the protocol for enrichment. This chapter provides guidelines and considerations for designing an affinity purification experiment, beginning with construction of a strain expressing a tagged subunit. Common biochemical methods for detecting protein, RNA, and DNA in Tetrahymena thermophila are also discussed.
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| http://dx.doi.org/10.1016/B978-0-12-385967-9.00012-8 | DOI Listing |
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