title: Quantification of fibronectin matrix assembly sites using a novel ELISA assay.
abstract: Binding of the N-terminus of fibronectin to assembly sites on the cell surface is an essential step in fibronectin fibrillogenesis. Fibronectin matrix assembly sites have customarily been quantified using an iodinated 70 kDa N-terminal fibronectin fragment. The 125I-70 K fragment is a less than ideal reagent because its preparation requires large amounts of plasma fibronectin and it has a fairly short shelf life. An additional limitation is that the cells responsible for binding the 125I-70 K cannot be quantified or identified directly but must be assessed in parallel cultures. To overcome these disadvantages, we developed an ELISA-based assay using a recombinant HA-tagged 70 K fragment. This assay allows for the simultaneous quantification and localization of matrix assembly sites on the surface of adherent cells.
authors.affiliation: Center for Cell Biology and Cancer Research, Albany Medical College, 47 New Scotland Avenue, Albany, New York 12208, USA.
authors.forename: Mingzhe
authors.initials: M
authors.lastname: Zheng
authors.forename: Anthony
authors.initials: A
authors.lastname: Ambesi
authors.forename: Lin
authors.initials: L
authors.lastname: Yu
authors.forename: Paula J
authors.initials: PJ
authors.lastname: McKeown-Longo
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005109:Extracellular Matrix / Q000378:metabolism; D005353:Fibronectins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D017434:Protein Structure, Tertiary; D019096:Vitronectin / Q000737:chemistry
chemical_list: D005353:Fibronectins; D019096:Vitronectin