ISSN:
1615-6102
Keywords:
Chlamydomonas
;
Cryofixation
;
Freeze-substitution
;
Immunocytochemistry
;
Nuclear proteins
;
Nucleus
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary We describe the preparation of monoclonal antibodies to nuclear antigens in the green alga,Chlamydomonas reinhardtii, and their localization at the light and electron microscope level. Supernatants from hybridomas were screened by the ELISA method and the four antibodies giving the strongest signal were subjected to further analysis. At the LM level immunogold silver staining was used on semi-thick resinless sections. We have examined at the EM level the distribution of these antigens by post-embedding immunocytochemical techniques on sections of conventionally fixed specimens compared to cryofixed and freeze-substituted ones. Enhanced ultrastructural preservation was observed in cells which were cryofixed, freeze-substituted and embedded at −35°C in Lowicryl K4M. Different preparative procedures involving cryofixation and substitution are described. Of the four antibodies three were localized under light and electron microscopy. All three were distributed in the interchromatin space. One of these antigens (QUL4D2, 54 kDa) is also found in the dense fibrillar component and fibrillar centers of the nucleolus.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01322289
