A preparation protocol for postembedding immunoelectron microscopy of Dictyostelium discoideum cells with monoclonal antibodies

Details

Serval ID
serval:BIB_9E59E247A333
Type
Article: article from journal or magazin.
Collection
Publications
Title
A preparation protocol for postembedding immunoelectron microscopy of Dictyostelium discoideum cells with monoclonal antibodies
Journal
Scanning Microscopy. Supplement
Author(s)
Humbel B.M., Biegelmann E.
ISSN
0891-7035
Publication state
Published
Issued date
1992
Volume
6
Number
3
Pages
817-825
Language
english
Abstract
On-section-labelling of Dictyostelium discoideum cells Poses severe Problems in retaining adequate morphology and antigenicity. Monoclonal antibodies are an essential tool in biochemistry and molecular biology because of their specificity and low background staining. Unfortunately their advantage, the recognition of only one distinct epitope, is often a handicap for immunoelectron microscopy. Resin embedding and steric hindrance of the gold-tagged secondary antibodies further reduce the efficiency in detecting antigens making the localization of less abundant antigens difficult if not impossible. A successful preparation protocol should retain morphology and antigenicity, allow the antibodies easy access to the antigen and use a detection system which visualizes as many of the primary antibodies as possible. Fixation of Dictyostelium cells with buffered formaldehyde and picric acid, cryosectioning and the use of ultra-small gold conjugates enabled us to label most of the antigens under investigation.
Keywords
CRYOULTRAMICROTOMY, ON-SECTION-LABELING, MONOCLONAL ANTIBODIES, ULTRA-SMALL GOLD CONJUGATES, SILVER ENHANCEMENT, AUTOMETALLOGRAPHY, DICTYOSTELIUM-DISCOIDEUM
Web of science
Create date
18/10/2012 13:31
Last modification date
20/08/2019 15:04
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