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Isolation, structural and immunohistochemical characterization of basement membrane heparan sulfate proteoglycan

Heparan sulfate proteoglycan (HSPG) was isolated from the mouse Engelbreth-Holm-Swarm (EHS) tumor. Polyclonal antibodies against the isolated HSPG were raised by two separate techniques, both producing highly reactive antisera. The antisera were found to be reactive against epithelial basement membrane of the mouse foot pad, glomerular basement membrane of the kidney and the EHS tumor matrix using the techniques of light microscopic immunoperoxidase on frozen sections and electron microscopic immunogold labelling on ultrathin Lowicryl K4M sections. At high magnification of the EHS tumor matrix, the gold particles were found over 5 nm wide sets of parallel lines, referred to as "double tracks" (Inoue et al., 1989). When HSPG was incubated in 50 mM Tris buffer, pH 7.4, at 35$ sp circ$C for either 5 min or 1 hr, a precipitate resulted which was made up of 5-6 nm wide sets of parallel lines, similar to double track structures. Together these studies demonstrate that HSPG of the EHS tumor matrix is made up of double track structures, and that in vitro incubation of HSPG produces double track structures in as short a time as 5 min.

Identiferoai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.61197
Date January 1992
CreatorsShima, Thomas Brent
PublisherMcGill University
Source SetsLibrary and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada
LanguageEnglish
Detected LanguageEnglish
TypeElectronic Thesis or Dissertation
Formatapplication/pdf
CoverageMaster of Science (Department of Anatomy.)
RightsAll items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated.
Relationalephsysno: 001265655, proquestno: AAIMM74880, Theses scanned by UMI/ProQuest.

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