RRC ID 77361
Author Oike Y, Hiraiwa H, Kawakatsu H, Nishikai M, Okinaka T, Suzuki T, Okada A, Yatani R, Sakakura T.
Title Isolation and characterization of human fibroblast tenascin. An extracellular matrix glycoprotein of interest for developmental studies.
Journal Int J Dev Biol
Abstract We have developed a biochemical method for purifying human tenascin from cultured fibroblasts or the culture medium. The method is a series of biochemical procedures including gel filtration, gelatin gel affinity chromatography and ion-exchange high performance liquid chromatography. The final preparation was identified as tenascin from its immunological cross-reactivity to antibody against chicken tenascin, strong hemagglutination activity which has been reported to be one of the biological functions of chicken tenascin, and from the electron microscopic study demonstrating a six-armed structure. Gel chromatography showed that intact human tenascin has an apparent molecular weight of over one million. Analysis of the purified tenascin with SDS-PAGE under reducing conditions demonstrated that tenascin consists of two kinds of subunits (250K and 190K). We established rat x mouse heterohybridoma cell lines which produce tenascin-specific antibodies. One monoclonal antibody (RCB1) was selected for immunohistochemical study and partially characterized. RCB1 bound native tenascin but not reduced and alkylated tenascin. Immunohistochemistry of normal and neoplastic tissues demonstrated that RCB1 bound the connective tissues surrounding the cancer nests and various normal tissues including interstitium of renal distal tubule, periosteum, endosteum, smooth muscles of digestive tract and media of arteries and arterioles.
Volume 34(2)
Pages 309-17
Published 1990-6-1
PMID 1696829
MeSH Antibodies, Monoclonal Cell Adhesion Molecules, Neuronal / immunology Cell Adhesion Molecules, Neuronal / isolation & purification* Cell Adhesion Molecules, Neuronal / pharmacology Cell Line Chromatography, Affinity Chromatography, Gel Chromatography, High Pressure Liquid Enzyme-Linked Immunosorbent Assay Fibroblasts / analysis* Hemagglutination Humans Hybridomas / immunology Immunoenzyme Techniques Microscopy, Electron Molecular Structure Molecular Weight Neoplasms / analysis Tenascin Tissue Distribution
IF 1.105
Human and Animal Cells P3-X63-Ag8.653(RCB0146)