A rapid and robust assay for detection of S-phase cell cycle progression in plant cells and tissues by using ethynyl deoxyuridine

Table 1 Comparison of experimental steps and durations for EdU-assay and BrdU-based immunolocalization

	EdU assay	BrdU immunolocalization
EdU or BrdU labeling	2 h*	2 h*
Fixation and washes	15 min + 15 min	15 min + 15 min
Cell wall digestion and washes	-	30 min + 15 min
Plasma membrane permeabilization and washes	-	30 min + 15 min
DNase I digestion and washes	-	30 min + 15 min
EdU click reaction or BrdU antibody incubations	30 min	3 h + 25 min + 1 h
Washing with DAPI and sample mounting	20 min	20 min
Total experimental duration	3 h 20 min	9 h 30 min

* Depending on the proliferation activity of a given cell type, shorter incubations may be sufficient or longer incubations may be necessary.

ISSN: 1746-4811