C6 NBD Ceramide

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Labeling of Golgi apparatus^[3]:
1. Rinse cells (on glass cover slips) by HMEM, then fix cells in glutaraldehyde or paraformaldehyde for 5-10 min (room temperature).
2. Wash fixed cells in a balanced salt solution (such as HCMF).
3. Transfer culture dishes containing the glass cover slips to an ice-water bath and incubate with freshly prepared NaBH₄ for 3×5 min (for glutaraldehyde-fixed cells).
4. Rinse cells several times (20-30 min) in cold HCMF, warm to room temperature, then incubate with C6 NBD-ceramide/BSA for 60 min.
5. Wash fixed cells by HCMF, and incubate with 10% fetal calf serum or 2 mg/mL BSA (30-90 min; room temperature) to back-exchange excess C6 NBD-ceramide from the preparation and enhance the staining of the Golgi apparatus.
6. The glass cover slips mounted on depression slides and observe by fluorescence microscope (Ex=466 nm, Em=536 nm).

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.