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Whole-Mount Immunolocalization Procedure for Plant Female Meiocytes
Santiago Valentín Galvan Gordillo1, Rocio Escobar-Guzman2, Daniel Rodriguez-Leal2,3
1Instituto de Biotecnología/UNAM, Cuernavaca, Morelos, Mexico.
Methods in Molecular Biology (Clifton, N.J.)
|October 5, 2019
Abstract:
Here we present an optimized protocol for immunolocalization of meiotic proteins during female meiosis in whole mount tissues. It ensures ovule morphology integrity and homogeneous reagent penetration. The method relies on paraformaldehyde tissue fixation, polyacrylamide embedding, tissue permeabilization, antibody incubation, counterstaining, and confocal microscopy analysis. This protocol has been used in diverse Arabidopsis ecotypes and in the legume Vigna unguiculata.