Fibronectin in cultured chick retinal pigment epithelial cells. 1. A specific antibody
Yamakawa, R.; Yoshimura, N.; Okada, M.; Kobayashi, H.; Matsumura, M.; Shirakawa, H.; Ogino, N.
Nippon Ganka Gakkai Zasshi 91(1): 80-86
1987
ISSN/ISBN: 0029-0203 PMID: 3296701 Document Number: 294585
To determine the production of fibronectin by cultured chick retinal pigment epithelial cells which are grown in the presence of fetal bovine serum, chick fibronectin-specific antisera, non-crossreactive to bovine fibronectin, was prepared. Fibronectin from fetal bovine serum and chick plasma were purified using Gelatin-Sepharose affinity chromatography and their purity was checked by SDS gel electrophoresis. Bovine fibronectin was coupled with CNBr activated Sepharose 4B column. Antisera for chick plasma fibronectin was applied to this column and non-crossing antisera was collected. Cross-reactivity was checked by immunoelectrophoretic blotting. Typical fibrillary net pattern of fibronectin was observed by indirect immunofluorescent technique when this specific antisera was reacted with cultured chick retinal pigment epithelial cells. Staining was abolished by absorption with the homologous antigen. Cultured chick retinal pigment epithelial cells were revealed to produce fibronectin.