BODIPY FL C5-Ceramide 

The Golgi apparatus is composed of flattened vesicles superimposed on each other by unit membranes. 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^[1]. BODIPY Fl C5-Ceramide is a Golgi-specific green fluorescent dye, which can visualise individual cells^[2]. Ex/Em= 505 nm/512 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.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

601.62

C₃₄H₅₄BF₂N₃O₃

133867-53-5

Solid

Brown to red

511

505

O=C(CCCCC1=CC=C2C=C3C(C)=CC(C)=[N]3[B+3]([F-])([N-]21)[F-])N[C@H]([C@@H](/C=C/CCCCCCCCCCCCC)O)CO

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

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

  [2]. Podgorny OV, et al. Isolation of single Chlamydia-infected cells using laser microdissection. J Microbiol Methods. 2015 Feb;109:123-8.  [Content Brief]