| Home > Publications database > Charakterisierung von Proteinen des Vertebraten-Photorezeptors mit Hilfe von monoklonalen Antikörpern |
| Book/Report | FZJ-2018-02572 |
1985
Kernforschungsanlage Jülich, Verlag
Jülich
Please use a persistent id in citations: http://hdl.handle.net/2128/18257
Report No.: Juel-2034
Abstract: Peripheral and soluble proteins of bovine rod outer segments were used as antigens for the production of hybridomas. Using radioimmunoassay light hybrid clones were selected with purified transducin (GBP) or proteinpreparations of cGMP dependent phosphodiesterase (PDE). The specificity of these hybridclones was determined by radioimmunoassay and western-blotting technique. For these experiments culture supernatants, ascites or isolated in mAb's were used. Three mAb's ($\gamma_{1}$-class) showed specific reactions with PDE from bovine rod outer segments. One of these antibodies inhibited the enzyme activity of PDE (hydrolysis of cGMP) between 40% and 50%. Proteinpreparations of evertebrate photoreceptorcells (Sepia officinalis) did not react with these three PDE-specific in mAb's. Five mAb's ($\mu$-class) crossreacted with several different peripheral and soluble proteins from bovine rod outer segments. These crossreactions were confirmed by several criterias. 1) Using enriched and purified protein preparations in radioimmunoassay. 2) Using extracts of peripheral and soluble proteins in the "Western-Blotting-technique". All proteins recognized by these mAb's interact with the photosensory membranes of bovine rod outer segments. Seven mAb"s were studied for their binding to SDS-denatured preparations of transducin (GBP)- and PDE-preparations using radioimmunoassay. Three different types of reactions were detected, and described.native andtypes
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