Published: Vol 2, Iss 19, Oct 5, 2012 DOI: 10.21769/BioProtoc.270 Views: 20682
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Abstract
This protocol describes measurement of direct protein-protein interactions by actin co-sedimentation assay. The actin co-sedimentation assay is a well-established technique and has been commonly used to demonstrate binding of proteins that interact directly with actin filaments (Ahrens et al., 2012; Mehta and Sibley, 2010; Schuler et al., 2005; Singh et al., 2011). We and others have previously shown that the damaged cell-recognition molecule C-type lectin 9A (Clec9A) recognises a conserved component within nucleated and non-nucleated cells that is exposed only when the cell membrane is damaged (Srivastava and Barber, 2008; Zhang et al., 2012). Here we use Clec9A as an example and present a detailed procedure for demonstrating the direct binding of purified recombinant Clec9A ectodomain to actin filaments.
Keywords: ActinMaterials and Reagents
Equipment
Procedure
Recipes
Acknowledgments
Jian-Guo Zhang was supported by Australian Government NHMRC Program Grants 461219 and 1016647. His work was also made possible through Victorian State Government Operational Infrastructure Support and Australian Government NHMRC IRIISS.
References
Article Information
Copyright
© 2012 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
Wong, W., Zhang, J., Baum, J., Lahoud, M. H. and Shortman, K. (2012). Co-sedimentation Assay for the Detection of Direct Binding to F-actin. Bio-protocol 2(19): e270. DOI: 10.21769/BioProtoc.270.
Category
Biochemistry > Protein > Interaction
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