Immunopurification of adenomatous polyposis coli (APC) proteins
- Author(s)
- Elliott, KL; Catimel, B; Church, NL; Coates, JL; Burgess, AW; Layton, MJ; Faux, MC;
- Details
- Publication Year 2013-10-25,Volume 6,Issue #1,Page 429
- Journal Title
- BMC Research Notes
- Publication Type
- Journal Article
- Abstract
- BACKGROUND: The adenomatous polyposis coli (APC) tumour suppressor gene encodes a 2843 residue (310 kDa) protein. APC is a multifunctional protein involved in the regulation of beta-catenin/Wnt signalling, cytoskeletal dynamics and cell adhesion. APC mutations occur in most colorectal cancers and typically result in truncation of the C-terminal half of the protein. RESULTS: In order to investigate the biophysical properties of APC, we have generated a set of monoclonal antibodies which enable purification of recombinant forms of APC. Here we describe the characterisation of these anti-APC monoclonal antibodies (APC-NT) that specifically recognise endogenous APC both in solution and in fixed cells. Full-length APC(1--2843) and cancer-associated, truncated APC proteins, APC(1--1638) and APC(1--1311) were produced in Sf9 insect cells. CONCLUSIONS: Recombinant APC proteins were purified using a two-step affinity approach using our APC-NT antibodies. The purification of APC proteins provides the basis for detailed structure/function analyses of full-length, cancer-truncated and endogenous forms of the protein.
- Publisher
- BioMed Central
- Research Division(s)
- Structural Biology
- Publisher's Version
- https://doi.org/10.1186/1756-0500-6-429
- Open Access at Publisher's Site
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- © 2013 Elliott et al.; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Creation Date: 2014-02-27 01:08:05