title: Immunopurification of adenomatous polyposis coli (APC) proteins.
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 β-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.
authors.forename: Kerryn L
authors.initials: KL
authors.lastname: Elliott
authors.forename: Bruno
authors.initials: B
authors.lastname: Catimel
authors.forename: Nicole L
authors.initials: NL
authors.lastname: Church
authors.forename: Janine L
authors.initials: JL
authors.lastname: Coates
authors.forename: Antony W
authors.initials: AW
authors.lastname: Burgess
authors.forename: Meredith J
authors.initials: MJ
authors.lastname: Layton
authors.affiliation: Ludwig Institute for Cancer Research-Parkville branch, Parkville, VIC 3050, Australia. faux@wehi.edu.au.
authors.forename: Maree C
authors.initials: MC
authors.lastname: Faux
mesh_terms: D025601:Adenomatous Polyposis Coli Protein / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D000941:Antigens / Q000008:administration & dosage / Q000737:chemistry; D016367:Baculoviridae / Q000235:genetics; D002846:Chromatography, Affinity / Q000379:methods*; D004285:Dogs; D015870:Gene Expression; D006801:Humans; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D061987:Sf9 Cells; D018411:Spodoptera
chemical_list: C555337:APC protein, human; D025601:Adenomatous Polyposis Coli Protein; D000911:Antibodies, Monoclonal; D000941:Antigens; D034741:RNA, Small Interfering; D011994:Recombinant Proteins