One for all--a highly efficient and versatile method for fluorescent immunostaining in fish embryos

For the detection and sub-cellular (co)-localization of proteins in the context of the tissue or organism immunostaining in whole mount preparations or on sections is still the best approach. So far, each antibody required its own fixation and antigen retrieval protocol so that optimizing immunostai...

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Bibliographic Details
Published in:PloS one 2011-05, Vol.6 (5), p.e19713-e19713
Main Authors: Inoue, Daigo, Wittbrodt, Joachim
Format: Article
Language:English
Subjects:
Animal genetic engineering
Animals
Antibodies
Antigens
Antigens - analysis
Biology
Cell morphology
Cytology
Danio rerio
Embryo, Nonmammalian - chemistry
Embryonic development
Embryos
Fishes
Fluorescence
Genetically modified animals
Genomes
Immunoglobulins
Immunohistochemistry - methods
Localization
Methods
Morphogenesis
Morphology
Proteins
Proteins - analysis
Stem cells
Transgenic fish
Zebrafish
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Summary:For the detection and sub-cellular (co)-localization of proteins in the context of the tissue or organism immunostaining in whole mount preparations or on sections is still the best approach. So far, each antibody required its own fixation and antigen retrieval protocol so that optimizing immunostaining turned out to be tedious and time consuming. Here we present a novel method to efficiently retrieve the antigen in a widely applicable standard protocol, facilitating fluorescent immunostaining of both cryosections and whole mount preparations in zebrafish (Danio rerio) and medaka (Oryzias latipes). Our method overcomes the loss of sections and damage of tissue and cell morphology, and allows parallel immunostaining in multiple colors, co-immunostaining with fluorescent proteins in transgenic fish lines and in combination with whole mount in situ hybridization.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0019713