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PMID: 3161899 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Heparan sulfate proteoglycans from mouse mammary epithelial cells: localization on the cell surface with a monoclonal antibody.

The Journal of cell biology ·Vol. 101 ·No. 3 ·1985-09-00 ·Pages 976-84

Jalkanen M, Nguyen H, Rapraeger A, Kurn N, Bernfield M

Abstract

Mouse mammary epithelial cells, of the normal murine mammary gland (NMuMG) cell line, bear a heparan sulfate-rich proteoglycan (HSPG) on their surfaces. A hybridoma (281-2) secreting a monoclonal antibody that recognizes this HSPG was produced by fusion of SP-2/0 myeloma cells with spleen cells from rats immunized with NMuMG cells. The 281-2 monoclonal antibody is directed against the core protein of the cell surface HSPG, as demonstrated by (a) recognition of the isolated proteoglycan but not its glycosaminoglycan chains, (b) co-localization of 281-2-specific antigen and radioactive cell surface HSPG on gradient polyacrylamide gel electrophoresis and on isopycnic centrifugation, and (c) abolition of immunofluorescent staining of the NMuMG cell surface by the intact, but not the protease-digested ectodomain of the cell surface HSPG. The antibody is specific for cell surface HSPG and does not recognize the HSPG that accumulates extracellularly beneath the basal cell surface. Therefore, the 281-2 antibody may be used to isolate the cell surface HSPG and to explore its distribution in tissues.

MeSH Terms
Aggrecans Animals Antibodies, Monoclonal/immunology Antibody Specificity Cell Line Chondroitin Sulfate Proteoglycans/immunology,metabolism Cytoplasm/immunology Epithelium Extracellular Matrix/immunology Extracellular Matrix Proteins Female Glycoproteins/immunology Glycosaminoglycans/immunology Heparan Sulfate Proteoglycans Heparitin Sulfate/immunology,metabolism Lectins, C-Type Mammary Glands, Animal Mice Proteoglycans/immunology
Chemicals
Acan protein, mouse Aggrecans Antibodies, Monoclonal Chondroitin Sulfate Proteoglycans Extracellular Matrix Proteins Glycoproteins Glycosaminoglycans Heparan Sulfate Proteoglycans Lectins, C-Type Proteoglycans Heparitin Sulfate
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Jalkanen M
Nguyen H
Rapraeger A
Kurn N
Bernfield M
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36 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.101.3.976
Published
1985-09-00
Pages
976-84
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2113719
Subset
IM
Grants
NCI NIH HHS · CA28735 · United States
NICHD NIH HHS · HD06763 · United States
NICHD NIH HHS · HD17146 · United States
External Links
PubMed source DOI: 10.1083/jcb.101.3.976 PMC Full Text View journal details
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