C6 NBD Galactosylceramide

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.