---
_id: '411'
abstract:
- lang: eng
text: Immunolocalization is a valuable tool for cell biology research that allows
to rapidly determine the localization and expression levels of endogenous proteins.
In plants, whole-mount in situ immunolocalization remains a challenging method,
especially in tissues protected by waxy layers and complex cell wall carbohydrates.
Here, we present a robust method for whole-mount in situ immunolocalization in
primary root meristems and lateral root primordia in Arabidopsis thaliana. For
good epitope preservation, fixation is done in an alkaline paraformaldehyde/glutaraldehyde
mixture. This fixative is suitable for detecting a wide range of proteins, including
integral transmembrane proteins and proteins peripherally attached to the plasma
membrane. From initiation until emergence from the primary root, lateral root
primordia are surrounded by several layers of differentiated tissues with a complex
cell wall composition that interferes with the efficient penetration of all buffers.
Therefore, immunolocalization in early lateral root primordia requires a modified
method, including a strong solvent treatment for removal of hydrophobic barriers
and a specific cocktail of cell wall-degrading enzymes. The presented method allows
for easy, reliable, and high-quality in situ detection of the subcellular localization
of endogenous proteins in primary and lateral root meristems without the need
of time-consuming crosses or making translational fusions to fluorescent proteins.
alternative_title:
- Methods in Molecular Biology
author:
- first_name: Michael
full_name: Karampelias, Michael
last_name: Karampelias
- first_name: Ricardo
full_name: Tejos, Ricardo
last_name: Tejos
- first_name: Jirí
full_name: Friml, Jirí
id: 4159519E-F248-11E8-B48F-1D18A9856A87
last_name: Friml
orcid: 0000-0002-8302-7596
- first_name: Steffen
full_name: Vanneste, Steffen
last_name: Vanneste
citation:
ama: 'Karampelias M, Tejos R, Friml J, Vanneste S. Optimized whole mount in situ
immunolocalization for Arabidopsis thaliana root meristems and lateral root primordia.
In: Ristova D, Barbez E, eds. Root Development. Methods and Protocols.
Vol 1761. MIMB. Springer; 2018:131-143. doi:10.1007/978-1-4939-7747-5_10'
apa: Karampelias, M., Tejos, R., Friml, J., & Vanneste, S. (2018). Optimized
whole mount in situ immunolocalization for Arabidopsis thaliana root meristems
and lateral root primordia. In D. Ristova & E. Barbez (Eds.), Root Development.
Methods and Protocols (Vol. 1761, pp. 131–143). Springer. https://doi.org/10.1007/978-1-4939-7747-5_10
chicago: Karampelias, Michael, Ricardo Tejos, Jiří Friml, and Steffen Vanneste.
“Optimized Whole Mount in Situ Immunolocalization for Arabidopsis Thaliana Root
Meristems and Lateral Root Primordia.” In Root Development. Methods and Protocols,
edited by Daniela Ristova and Elke Barbez, 1761:131–43. MIMB. Springer, 2018.
https://doi.org/10.1007/978-1-4939-7747-5_10.
ieee: M. Karampelias, R. Tejos, J. Friml, and S. Vanneste, “Optimized whole mount
in situ immunolocalization for Arabidopsis thaliana root meristems and lateral
root primordia,” in Root Development. Methods and Protocols, vol. 1761,
D. Ristova and E. Barbez, Eds. Springer, 2018, pp. 131–143.
ista: 'Karampelias M, Tejos R, Friml J, Vanneste S. 2018.Optimized whole mount in
situ immunolocalization for Arabidopsis thaliana root meristems and lateral root
primordia. In: Root Development. Methods and Protocols. Methods in Molecular Biology,
vol. 1761, 131–143.'
mla: Karampelias, Michael, et al. “Optimized Whole Mount in Situ Immunolocalization
for Arabidopsis Thaliana Root Meristems and Lateral Root Primordia.” Root
Development. Methods and Protocols, edited by Daniela Ristova and Elke Barbez,
vol. 1761, Springer, 2018, pp. 131–43, doi:10.1007/978-1-4939-7747-5_10.
short: M. Karampelias, R. Tejos, J. Friml, S. Vanneste, in:, D. Ristova, E. Barbez
(Eds.), Root Development. Methods and Protocols, Springer, 2018, pp. 131–143.
date_created: 2018-12-11T11:46:20Z
date_published: 2018-03-11T00:00:00Z
date_updated: 2021-01-12T07:54:34Z
day: '11'
department:
- _id: JiFr
doi: 10.1007/978-1-4939-7747-5_10
editor:
- first_name: Daniela
full_name: Ristova, Daniela
last_name: Ristova
- first_name: Elke
full_name: Barbez, Elke
last_name: Barbez
intvolume: ' 1761'
language:
- iso: eng
month: '03'
oa_version: None
page: 131 - 143
publication: Root Development. Methods and Protocols
publication_status: published
publisher: Springer
publist_id: '7418'
quality_controlled: '1'
scopus_import: 1
series_title: MIMB
status: public
title: Optimized whole mount in situ immunolocalization for Arabidopsis thaliana root
meristems and lateral root primordia
type: book_chapter
user_id: 4435EBFC-F248-11E8-B48F-1D18A9856A87
volume: 1761
year: '2018'
...