An Optimized Whole-Mount Immunofluorescence Method for Shoot Apices

Curr Protoc. 2021 Apr;1(4):e101. doi: 10.1002/cpz1.101.

Abstract

The localization of a protein provides important information about its biological functions. The visualization of proteins by immunofluorescence has become an essential approach in cell biology. Here, we describe an easy-to-follow immunofluorescence protocol to localize proteins in whole-mount tissues of maize (Zea mays) and Arabidopsis. We present the whole-mount immunofluorescence procedure using maize ear primordia and Arabidopsis inflorescence apices as examples, followed by tips and suggestions for each step. In addition, we provide a supporting protocol to describe the use of an ImageJ plug-in to analyze colocalization. This protocol has been optimized to observe proteins in 2-5 mm maize ear primordia or in developing Arabidopsis inflorescence apices; however, it can be used as a reference to perform whole-mount immunofluorescence in other plant tissues and species. © 2021 Wiley Periodicals LLC. Basic Protocol: Whole-mount immunofluorescence for maize and Arabidopsis shoot apices Support Protocol: Measure colocalization by JACoP plugin in ImageJ.

Keywords: Arabidopsis; immunofluorescence; maize; protein localization; whole-mount.

MeSH terms

  • Arabidopsis Proteins*
  • Arabidopsis*
  • Fluorescent Antibody Technique
  • Inflorescence
  • Zea mays

Substances

  • Arabidopsis Proteins