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Nat Protoc. 2006;1(1):98-103.

Immunocytochemical techniques for whole-mount in situ protein localization in plants.

Author information

1
Center for Plant Molecular Biology, Tübingen University, 72076 Tübingen, Germany.

Abstract

As the field of plant molecular biology is swiftly advancing, a need has been created for methods that allow rapid and reliable in situ localization of proteins in plant cells. Here we describe a whole-mount 'immunolocalization' technique for various plant tissues, including roots, hypocotyls, cotyledons, young primary leaves and embryos of Arabidopsis thaliana and other species. The detailed protocol, recommended controls and troubleshooting are presented, along with examples of applications. The protocol consists of five main procedures: tissue fixation, tissue permeation, blocking, primary and secondary antibody incubation. Notably, the first procedure (tissue fixation) includes several steps (4-12) that are absolutely necessary for protein localization in hypocotyls, cotyledons and young primary leaves but should be omitted for other tissues. The protocol is usually done in 3 days, but could also be completed in 2 days.

PMID:
17406218
DOI:
10.1038/nprot.2006.15
[Indexed for MEDLINE]

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