@inbook{732b2351a84349cab897155d67af9022,
title = "Using Tandem Affinity Purification to Identify Circadian Clock Protein Complexes from Arabidopsis",
abstract = "Identification of protein-protein interactions is an effective method of elucidating new roles for circadian clock-associated proteins that can expand beyond the information collected from transcriptional studies and genetic screens. Tandem affinity purification coupled with liquid chromatography mass spectrometry (APMS) utilizes epitope-tagged versions of your protein of interest to co-precipitate direct and indirect protein partners. Here, we provide a protocol and suggestions for proper design of 6x-His-3x-FLAG-tagged clock proteins and isolation of protein-protein interactions using two immunoprecipitation steps for increased specificity.",
keywords = "Affinity purification, Circadian clock, Mass spectrometry, Protein-protein interactions",
author = "Sorkin, {Maria L.} and Nusinow, {Dmitri A.}",
note = "Funding Information: M. L. Sorkin is supported by NSF GRF award number DGE-1745038. This protocol is based upon material analyzed using an Orbitrap Fusion Lumos LC-MS/MS supported by the National Science Foundation under Grant No. DBI-1827534. Publisher Copyright: {\textcopyright} 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2022",
doi = "10.1007/978-1-0716-1912-4_15",
language = "English",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "189--203",
booktitle = "Methods in Molecular Biology",
}