Immunoelectron Microscopy of Cryofixed Freeze-Substituted Yeast Saccharomyces cerevisiae

0503597 - ÚMG 2019 RIV US eng J - Článek v odborném periodiku
Fišerová, Jindřiška - Richardson, C. - Goldberg, M.W.
Immunoelectron Microscopy of Cryofixed Freeze-Substituted Yeast Saccharomyces cerevisiae.
Methods in Molecular Biology. Methods in Molecular Biology. Roč. 1474, January (2016), s. 243-258. ISSN 1064-3745
Institucionální podpora: RVO:68378050
Klíčová slova: Yeast * Nuclear * Transport * Transmission electron microscopy * Freeze substitution * Lowicryl * Immunostaining
Obor OECD: Biochemistry and molecular biology

Immunolabeling electron microscopy is a challenging technique with demands for perfect ultrastructural and antigen preservation. High-pressure freezing offers an excellent way to fix cellular structure. However, its use for immunolabeling has remained limited because of the low frequency of labeling due to loss of protein antigenicity or accessibility. Here we present a protocol for immunogold labeling of the yeast Saccharomyces cerevisiae that gives specific and multiple labeling while keeping the finest structural details. We use the protocol to reveal the organization of individual nuclear pore complex proteins and the position of transport factors in the yeast Saccharomyces cerevisiae in relation to actual transport events.
Trvalý link: http://hdl.handle.net/11104/0295408