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CAS:86701-10-2
Appearance:Soild
Storage:Powder : -20℃, 2 years;In solvent(mother liquid): -20℃, 1 month; -80℃, 6 months
Purity:≥98%
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NBD C6-Ceramide is a fluorescent sphingolipid analogue that can be used to study the mechanism of sphingolipid transport and metabolism. It can also be used to selectively stain the Golgi apparatus in living and fixed cells. Environmentally sensitive NBD dyes fluoresce weakly in water, but fluorescence increases in aprotic solvents and other non-polar environments.
How to use (for reference only)
Live cell staining
(1) The cells were cultured on sterile cover slides.
(2) When the cells are cultured to a suitable density, remove the cover glass from the medium and rinse the cover glass with a suitable buffer (such as serum-free medium, HBSS, HEPES or PBS).
(3) Add 100μL dye solution (5-10 μM) to a corner of the cover glass, gently shake to make the dye evenly cover all cells, and incubate at room temperature for 30 min.
(4) Blot the dyeing solution, clean the cover glass with fresh medium pre-cooled at 4 ℃ for 2 to 3 times, then cover all cells with fresh medium, and incubate at 37 ℃ for 30 min.
(5) After cleaning with fresh medium, observe with microscope.
Fixed cell staining
(1) The cells were cultured on sterile cover slides.
(2) When the cells are cultured to a suitable density, remove the cover glass from the medium and rinse the cover glass with a suitable buffer (such as serum-free medium, HBSS, HEPES or PBS). Use 4% paraformaldehyde to fix at room temperature for 5 to 10 minutes.
(3) Add 100μL of dye solution (5-10 μM) to one corner of the cover glass, gently shake the dye to evenly cover all cells, and incubate at room temperature for 30min.
(4) Blot the dyeing solution, clean the cover glass with the same buffer solution, and then incubate it with 10% FBS or 2 mg/mL BSA at room temperature for 30-90 min to improve the Golgi staining.
(5) After cleaning with the same buffer, observe with a microscope.
Matters needing attention
1. Fluorescent dyes have quenching problems, please try to avoid light to slow down fluorescence quenching.
2. If the suspension cells are stained with Golgi apparatus, it is recommended to stain at 2x106 cells /mL.
3. For your safety and health, please wear a lab coat and disposable gloves.
