BODIPY TR Ceramide

BODIPY TR Ceramide (Golgi-Red Tracke) is a Golgi-specific fluorescent dye, which can visualise individual cells[1]. Ex/Em=589 nm/616 nm. The flattened vesicles are round with expanded and perforated edges. The Golgi fluorescent probe is a BODIPY-labeled ceramide derivative, the synthesis of which occurs in the endoplasmic reticulum and can then be transported to the Golgi via ceramide transport protein (CERT) or vesicular translocation, allowing specific labeling of the dye. BODIPY TR Ceramide (Golgi-Red Tracke) is a Golgi-specific fluorescent dye, which can visualise individual cells. Ex/Em=589 nm/616 nm.

General Protocol
1 Preparation of Golgi working solution
1.1 Preparation of the stock solution
Dissolve Golgi in DMSO to obtain 5 mM of Golgi.
Note: It is recommended to store the stock solution at -20°C -80°C away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of Golgi working solution
Dilute the stock solution in HBSS to obtain 1-10 μM of Golgi working solution.
Note: Please adjust the concentration of Golgi working solution according to the actual situation.2 Cell staining
2.1 Suspension cells (6-well plate)
a.Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.The cell density is 1×10⁶/mL.
b.Add 1 mL of working solution, and then incubate at room temperature for 20-30 minutes.
c.Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
d.Wash twice with PBS, 5 minutes each time.
e.Resuspend cells with serum-free cell culture medium or PBS.Observation by fluorescence microscopy or flow cytometry.
2.2 Adherent cells
a.Culture adherent cells on sterile coverslips.
b.Remove the coverslip from the medium and aspirate excess medium.
c.Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 20-30 minutes.
d.Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or flow cytometry.

Storage
-20℃ 1 year.Protect from light.

Precautions
1. It is recommended to store the stock solution at -20℃ or -80℃ away from light and avoid repetitive freeze-thaw cycles.
2. Please adjust the concentration of Golgi 1-43 working solution according to the actual situation.
3. when the cell culture solution is removed with poor results, the cells can be washed with an appropriate amount of Hanks balanced salt solution.
4. This product is for R&D use only, not for drug, household, or other uses.
5. For your safety and health, please wear a lab coat and disposable gloves to operate.

References:
[1]. Alamudi SH, et al. Development of background-free tame fluorescent probes for intracellular live cell imaging. Nat Commun. 2016 Jun 20;7:11964.