A rapid and robust assay for detection of S | 合法醫療器材資訊網

由EKotogány著作·2010·被引用162次—ProgressinplantcellcycleresearchishighlydependentonreliablemethodsfordetectionofcellsreplicatingDNA.FrequencyofS-phasecells ...

To compare EdU assay with BrdU immunodetection, Arabidopsis thaliana sp. Columbia suspension culture cells were incubated with 10 μM EdU for 2 hrs and fixed with detergent-containing formaldehyde. Since EdU detection does not rely on antibodies and the molecular size of the chemicals used during detection is small compared to the size of the antibody molecules, cells were directly incubated with the detection cocktail without performing cell wall digestion. The detection cocktail is composed of an azide-conjugated fluorochrome (such as Alexa Fluor 488) and copper (I) as the catalyzer of the click reaction [13]. Stained nuclei with high signal-to-noise ratio were easily detectable under the microscope (Figure 1A). Omission of cell wall digestion and DNA denaturation/digestion steps resulted in better preservation of cellular structures judged by DAPI (4,6-diami...