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Title

Advances in multilabeling for TEM and CLSM using primary antibodies derived from the same species. Application of Fab fragments to co-localize olive pollen allergens

AuthorsMorales, Sonia; Rodríguez García, María I.; Alché Ramírez, Juan de Dios
Keywordsallergen
antibodies
Fab fragments
immunofluorescence
immunogold
olive
pollen
multilabeling
Issue DateDec-2010
PublisherFormatex
CitationMicroscopy: Science, Technology, Applications and Education
AbstractThe cell distribution of multiple antigens can be simultaneously revealed by fluorescence and confocal laser scanning microscopy (FM, CLSM) after using antibodies (Abs) conjugated to fluorophores with differential spectral characteristics. The same multiple localization can be carried out at TEM (Transmission Electron Microscopy) by using gold particles of different sizes. However, multilabeling studies often need to use primary Abs derived from the same host species. In these cases, interference between different rounds of Abs must be prevented. Methods available to tackle this need are reviewed, focusing particularly on the use of Fab fragments. Protocols regarding multiple localization of olive pollen targets (Ole e 1, Ole e 2, Ole e 9 and actin) by CLSM and TEM are provided. They relay on the use of primary Abs raised in the same host species (rabbit) and the application of Fab fragments.
DescriptionPages of the mentioned chapter: 607-613 (both included) Publisher: Formatex Research Center
URIhttp://hdl.handle.net/10261/32446
ISBNVolume 1 ISBN (13): 978-84-614-6189-9
Appears in Collections:(EEZ) Libros y partes de libros
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