Cell Navigator® TMR Ceramide Golgi Staining Kit *Red Fluorescence*
![<strong>The representative fluorescence image of GGR169 Ceramide Golgi Staining in HeLa cells.</strong> Cells were stained with 100 µL of GGR169-ceramide working solution with Hoechst 33342 at 37 °C for 20 minutes. An intensely fluorescent thread like structure, partially surround the nucleus, is identified as the Golgi apparatus.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-navigator-tmr-ceramide-golgi-staining-kit-red-fluorescence%2Ffigure-for-cell-navigator-tmr-ceramide-golgi-staining-kit-red-fluorescence_iqUxe.jpg&w=640&q=75)
![<strong>The representative fluorescence image of GGR169 Ceramide Golgi Staining in HeLa cells.</strong> Cells were stained with 100 µL of GGR169-ceramide working solution with Hoechst 33342 at 37 °C for 20 minutes. An intensely fluorescent thread like structure, partially surround the nucleus, is identified as the Golgi apparatus.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-navigator-tmr-ceramide-golgi-staining-kit-red-fluorescence%2Ffigure-for-cell-navigator-tmr-ceramide-golgi-staining-kit-red-fluorescence_iqUxe.jpg&w=640&q=75)
![<strong>The representative fluorescence image of GGR169 Ceramide Golgi Staining in HeLa cells.</strong> Cells were stained with 100 µL of GGR169-ceramide working solution with Hoechst 33342 at 37 °C for 20 minutes. An intensely fluorescent thread like structure, partially surround the nucleus, is identified as the Golgi apparatus.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-navigator-tmr-ceramide-golgi-staining-kit-red-fluorescence%2Ffigure-for-cell-navigator-tmr-ceramide-golgi-staining-kit-red-fluorescence_iqUxe.jpg&w=128&q=25)
![<strong>The representative fluorescence images of GGR169 Ceramide Golgi Staining in HeLa cells in a pH dependent manner.</strong> HeLa cells were stained with 100 µL of GGR169-ceramide working solution at 37 °C for 20 minutes. Images were acquired using a fluorescence microscope equipped with Cy3 filter set before and after the addition of 30 mM of NH<sub>4</sub>Cl solution. A reduction in fluorescence intensity was observed after the addition of NH<sub>4</sub>Cl.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-navigator-tmr-ceramide-golgi-staining-kit-red-fluorescence%2Ffigure-for-cell-navigator-tmr-ceramide-golgi-staining-kit-red-fluorescence_xj9jW.jpg&w=128&q=25)
AT A GLANCE
- Treat cells as desired
- Add GGR169-ceramide working solution and incubate at room temperature or 37 °C for 15∼30 minutes
- Replace with the Staining Buffer
- Observe under microscope using Cy3 filter set
Thaw all the components at room temperature before starting the experiment.
CELL PREPARATION
For guidelines on cell sample preparation, please visit https://www.aatbio.com/resources/guides/cell-sample-preparation.html
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles
Add 100 µL of DMSO (Component C) to GGR-ceramide (Component A) to make GGR169-ceramide stock solution (100X).
Note Store the unused GGR-ceramide stock solution at -20 °C in single use aliquots to avoid freeze thaw cycles.
PREPARATION OF WORKING SOLUTION
Add 10 µL of GGR169-ceramide stock solution (100X) to 990 µL of Staining Buffer (Component B) to make GGR169-ceramide working solution.
Optional: Add 10 µL of Hoechst 33342 (Component D) to 1 mL GGR169-ceramide working solution for nuclear stain. Observe under fluorescence microscope with DAPI filter set.
SAMPLE EXPERIMENTAL PROTOCOL
Following protocol should be used for the guidelines and can be modified as per requirements.
- Plate and treat cells as desired.
Remove the cell culture medium. Optional: Cells can be washed with buffer of your choice.
Add 100 µL of GGR169-ceramide working solution directly into cell culture medium.
Incubate at room temperature or 37 °C for 15∼30 minutes.
Remove the GGR169-ceramide working solution and wash once with DPBS or buffer of your choice.
Add 100 µL/well of Staining Buffer (Component B).
Observe under a fluorescence microscope with Cy3 filter set.
Name | Excitation (nm) | Emission (nm) |
Cell Navigator® NBD Ceramide Golgi Staining Kit *Green Fluorescence* | 467 | 538 |
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