C6 NBD Galactosylceramide [170212-26-7]

C6 NBD galactosylceramide is an active derivative of galactosylceramide that is tagged with fluorescent C6 nitrobenzoxadiazole (C6 NBD). C6 NBD galactosylceramide can be used as a substrate for neutral β-glycosylceramidase (GCase) to study intracellular localization and metabolism of galactosylceramide (Ex=nm, Em=525)^[1].

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Transcytosis of Exogenous C6-NBD-GaICer after Endocytosis^[2]
1. Add C6 NBD Galactosylceramide at 10 °C.
2. Rinse the cells three times with ice-cold HBSS buffer, then incubate in HBSS buffer at 37 °C to allow endocytosis.
3. After 10 min, remove the remaining probe on the cell surface by washing with BSA solution at 10 °C.
4. One set of filters is used for lipid analysis to quantify endocytosis efficiency; another set of filters is further incubated in HBSS buffer containing BSA at 37 °C for 0.5 or 1 h to detect the redistribution of intracellular C6 NBD Galactosylceramide on the cell surface.
5. After incubation, wash with BSA solution at 10 °C; then extract NBD lipids from the culture medium and cells using a chloroform/methanol system, separate by thin-layer chromatography (TLC), and perform quantitative analysis.

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

• [1]. J W Kok, et al. Sorting of sphingolipids in the endocytic pathway of HT29 cells. J Cell Biol. 1991 Jul;114(2):231-9.  [Content Brief]

  [2]. I van Genderen, et al. Differential targeting of glucosylceramide and galactosylceramide analogues after synthesis but not during transcytosis in Madin-Darby canine kidney cells. J Cell Biol. 1995 Nov;131(3):645-54.  [Content Brief]