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Phloem pp 109-122 | Cite as

Immunohistochemical Localization of Proteins in the Phloem: Problems and Solutions

  • Christina KühnEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2014)

Abstract

Immunolocalization of proteins in differentiated phloem cells is a challenging task given their special anatomy, organellar infrastructure, and the phloem tissue’s heterogeneity. Incorporation of specific wall components in the thickened cell walls of phloem cells is often the source of unspecific labeling, leading to erroneous localization. Therefore, special care is required regarding generation and purification of specific antibodies. In addition, tissue preservation of phloem cells, which contain a high osmotic pressure in their functional state, is a very challenging task prone to various pitfalls. This chapter provides practical advice for cautious tissue preparation and antibody purification. Furthermore, methods that can be used to verify immunohistochemical localization data, such as promoter-reporter studies or activity tests, are discussed. Such confirmation experiments are essential for unambiguous determination of protein location in cells of the phloem.

Key words

Phloem transport Carbon allocation Sucrose transporter Phloem proteins Immunolocalization Subcellular localization 

Notes

Acknowledgments

Critical proof-reading by Alexander Schulz, Johannes Liesche and Pawel Brzezowski was greatly appreciated.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Plant Physiology, Institute of BiologyHumboldt University BerlinBerlinGermany

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