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Immunoelectron Microscopy

Methods and Protocols
Series: Methods in Molecular Biology, Vol. 657
Schwartzbach, Steven D.; Osafune, Tetsuaki (Eds.)
1st Edition., 2010, XI, 352 p. 82 illus., 41 in color., Hardcover
ISBN: 978-1-60761-782-2

Due: July 29, 2010
152,01$

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About this book

Immunoelectron microscopy is a key technique that bridges the information gap between biochemistry, molecular biology, and ultrastructural studies placing macromolecular functions within a cellular context. In Immunoelectron Microscopy: Methods and Protocols, expert researchers combine the tools of the molecular biologist with those of the microscopist. From the molecular biology toolbox, this volume presents methods for antigen production by protein expression in bacterial cells, methods for epitope tagged protein expression in plant and animal cells allowing protein localization in the absence of protein specific antibodies as well as methods for the production of anti-peptide, monoclonal, and polyclonal antibodies. From the microscopy toolbox, sample preparation methods for cells, plant, and animal tissue are presented. Both cryo-methods, which have the advantage of retaining protein antigenicity at the expense of ultrastructural integrity, as well as chemical fixation methods that maintain structural integrity while sacrificing protein antigenicity have been included, with chapters examining various aspects of immunogold labeling. Written in the highly successful Methods in Molecular Biology? series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and essential, Immunoelectron Microscopy: Methods and Protocols seeks to facilitate an increased understanding of structure function relationships.
Content Level " Professional/practitioner
Keywords " Fixation protocols - Macromolecular functions - Pre- and post-embedding immunogold labeling - Structure function relationships
Related subjects " Cell Biology - Immunology

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Formation of "giant chloroplast" during the cell cycle of Euglena gracilis Z in synchronized culture
Osafune,T,,Tanaka,K. and Ehara,T.
Cytorogia, Vol.74,1,(2010)

In the cell cycle of Euglena gracilis Z in synchronized cultures, chloroplasts temporarily conjoin to form a single giant structure at the 14th hour after the onset of the light period, called "giant chloroplast" (upper in right and bottom), which surrounds the nucleus making connections or close contacts at several sites. The upper left shows the 10th hour after the onset of the light period. The cell contained 11 chloroplasts and chloroplasts were round or oval in shapes and dispersed in the cytoplasm. Chloroplast nucleoids in the "giant chloroplast", observed under a fluorescence microscope after staining with DAPI, the DNA fluorochrome, become stringy and some tips of the string appeared to come into close proximity to the site of connection with the nucleus (upper in right). Bar (upper, bottom: 5mm).

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Ehara,T.,Osafune,T.,Hase, E.:
Interactions between the nucleus and cytoplasmic organelles during the cell cycle of Euglena gracilis in synchronous cultures.

Exp. Cell Res.190(1)104-112, (1990)
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Schwartzbach,S.D., Osafune, T., Löffelhardt, W.:
Protein import into cyanelles and complex chloroplasts.

Plant Molecular Biology. 382(2)263-274 (1998)
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Osafune,T.,Ehara,T.,Sumida,S.,Hase,E., Schiff,J.A.:
Light-independent processes in the formation of thylakoids and pyrenoids in proplastids of dark-grown cells of Euglena gracilis.

J. Electron Microscopy. 39(4)245-253, (1990)
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A three-dimensional computer generated model of synchronously dividing Euglena gracilis Z cell at light/dark cycle constructed based on serial sections.

Cytologia. 67,
(1)1-2, (2000)
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Osafune,T.,Sumida,S.,Ehara,T.,Hase,E.:
Three-dimensional distribution of ribulose-1,5-bisphosphate carboxylase oxygenase in chloroplasts of actively photosynthesizing cell of Euglena gracilis.

J. Electron Microscopy,38(5)399-402, (1989)

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�@copyright (c) 2002 Tetsuaki Osafune All rights reserved