The fluorescent properties of IB were utilized for localization analyses. IB is excited at 568 and emits fluorescence at >650. ER-tracker Green was purchased from Thermo-Fisher (E34250) and Golgi-ID and Lyso-ID were purchased from Enzo (ENZ-51028-K100 and ENZ-51034-0100). These dyes were chosen because they have no spectral overlap with IB. Hoechst stain provided in the Enzo kits was used for nuclear counterstain. Staining was performed according to manufacturer’s protocols. Cells were concurrently loaded with 150 nM IB. High resolution live-cell imaging was performed on the DeltaVision OMX (GE Healthcare). Post-acquisition data analysis was done using Fiji software (Fiji.sc).
Golgi id
Manufactured by Enzo Life Sciences
Sourced in United Kingdom
Golgi-ID is a fluorescent labeling reagent that specifically stains the Golgi apparatus in live cells. It provides a simple and reliable method for visualizing the Golgi complex in a variety of cell types.
Automatically generated - may contain errors
2 protocols using golgi id
1
Fluorescent Dyes for Organelle Localization
2
Golgi-ID Staining of A549 Cells
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Staining using Golgi-ID (Enzo Life Sciences, Exeter, UK) was achieved according to the manufacturer’s instructions. Briefly, A549 cells were plated as previously described. Cells were incubated overnight and washed with 100 µL of assay solution before the addition of a 200 µL of Golgi-ID stain (1 in 100 dilution of provided stock solution) per well. Cells were then incubated at 4 °C for 30 min before removal of the Golgi-ID stain and washing of the cells with 100 µL assay buffer. A final 1 mL of DMEM was added to the cells and they were incubated for 30 min at 37 °C in a 5% CO2 incubator with ~95% humidity before use in the ricin assays.
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