Affinity Purification of Protein Complexes in C. elegans
Zanin E, Dumont J, Gassmann R, Cheeseman I, Maddox P, Bahmanyar S, Carvalho A, Niessen S, Yates JR, Oegema K, Desai A (2011)
Publication Type: Book chapter / Article in edited volumes
Publication year: 2011
Journal
Publisher: Academic Press Inc.
Edited Volumes: Methods in Cell Biology
Series: Methods in Cell Biology
Book Volume: 106
Pages Range: 289-322
DOI: 10.1016/B978-0-12-544172-8.00011-6
Abstract
C. elegans is a powerful metazoan model system to address fundamental questions in cell and developmental biology. Research in C. elegans has traditionally focused on genetic, physiological, and cell biological approaches. However, C. elegans is also a facile system for biochemistry: worms are easy to grow in large quantities, the functionality of tagged fusion proteins can be assessed using mutants or RNAi, and the relevance of putative interaction partners can be rapidly tested in vivo. Combining biochemistry with function-based genetic and RNA interference screens can rapidly accelerate the delineation of protein networks and pathways in diverse contexts. In this chapter, we focus on two strategies to identify protein-protein interactions: single-step immunoprecipitation and tandem affinity purification. We describe methods for growth of worms in large-scale liquid culture, preparation of worm and embryo extracts, immunoprecipitation, and tandem affinity purification. In addition, we describe methods to test specificity of antibodies, strategies for optimizing starting material, and approaches to distinguish specific from non-specific interactions. © 2011 Elsevier Inc.
Authors with CRIS profile
Additional Organisation(s)
Involved external institutions
University of California, San Diego
United States (USA) (US)
Université de Montréal
Canada (CA)
Scripps Research Institute (TSRI)
United States (USA) (US)
Institut Curie
France (FR)
Massachusetts Institute of Technology (MIT)
United States (USA) (US)
United States (USA) (US)
Université de Montréal
Canada (CA)
Scripps Research Institute (TSRI)
United States (USA) (US)
Institut Curie
France (FR)
Massachusetts Institute of Technology (MIT)
United States (USA) (US)
How to cite
APA:
Zanin, E., Dumont, J., Gassmann, R., Cheeseman, I., Maddox, P., Bahmanyar, S.,... Desai, A. (2011). Affinity Purification of Protein Complexes in C. elegans. In Methods in Cell Biology. (pp. 289-322). Academic Press Inc..
MLA:
Zanin, Esther, et al. "Affinity Purification of Protein Complexes in C. elegans." Methods in Cell Biology. Academic Press Inc., 2011. 289-322.
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