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PMID: 2188187 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

Antigenic expression of neuron-associated class III beta-tubulin isotype (h beta 4) and microtubule-associated protein 2 (MAP2) by the human retinoblastoma cell line WERI-Rb1. A comparative immunoblot and immunocytochemical study.

Ophthalmic research ·Vol. 22 ·No. 1 ·1990-00-00 ·Pages 57-66

Gass P, Frankfurter A, Katsetos CD, Herman MM, Donoso LA, Rubinstein LJ

Abstract

The antigenic expression of two neuron-associated microtubule proteins, class III beta-tubulin isotype (h beta 4) and microtubule-associated protein 2 (MAP2), was evaluated in a comparative immunoblot and immunocytochemical study of the human retinoblastoma cell line WERI-Rb1 maintained for up to 30 days in three different in vitro conditions. Western blots were performed on whole sodium dodecyl sulfate extracts of cells grown in floating suspensions, on Gelfoam matrices and on coverslips. Immunoperoxidase histochemistry was performed on matrix cultures. Immunoblotting demonstrated that h beta 4 and MAP2 were present under all culture conditions. By immunocytochemistry, staining of cytologically undifferentiated cells with anti-h beta 4 and anti-MAP2 monoclonal antibodies was found on Gelfoam matrix explants. In contrast, glial fibrillary acidic protein was not detected by either immunoblots or immunocytochemistry. These findings are in keeping with the solely neuroblastic nature of this line and provide no evidence for its divergent (i.e. neuronal and glial) differentiation capacity.

MeSH Terms
Antibodies, Monoclonal Blotting, Western Electrophoresis, Polyacrylamide Gel Eye Neoplasms/metabolism Glial Fibrillary Acidic Protein/metabolism Humans Immunoenzyme Techniques Microtubule-Associated Proteins/biosynthesis Retina/metabolism Retinoblastoma/metabolism Tubulin/metabolism Tumor Cells, Cultured/metabolism
Chemicals
Antibodies, Monoclonal Glial Fibrillary Acidic Protein Microtubule-Associated Proteins Tubulin
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Gass P
Department of Pathology, University of Virginia School of Medicine, Charlottesville.
Frankfurter A
Katsetos C D
Herman M M
Donoso L A
Rubinstein L J
Article Info
Journal
Ophthalmic research
Abbr.
Ophthalmic Res
ISSN
0030-3747
Published
1990-00-00
Pages
57-66
Language
English
Region
Switzerland
NLM ID
0267442
Subset
IM
Grants
NCI NIH HHS · CA31271 · United States
NEI NIH HHS · EY7737 · United States
NINDS NIH HHS · NS21142 · United States
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