Bioactivity | 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]. Storage: protect from light. |
Invitro | C6-NBD-glucosylceramide (4 μM) is transported to the Golgi apparatus in HT29 cells[1].Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)[2].Transcytosis of Exogenous C6-NBD-GaICer after Endocytosis: 1. C6-NBD-glucosylceramide is inserted at 10°C. 2. Cells are rinsed three times with cold HBSS' and incubated at 37°C in HBSS' to allow endocytosis. 3. After 10 min, the probe remaining on the cell surface is removed by two (apical) or three (basal) BSA washes for 20 min at 10°C. 4. One set of filters is used for lipid analysis to quantitate endocytosis. A second set of filters is further incubated for 0.5 or 1 h at 37°C in HBSS' + BSA, to assay for reappearance of intracellular C6-NBD-glucosylceramide on either cell surface. 5. The incubations are followed by a 10°C BSA wash, after which the NBD lipids from the combined apical media, basal media, and the cells were extracted into chloroform/methanol, analyzed by TLC, and quantitated. |
Name | C6 NBD Galactosylceramide |
CAS | 170212-26-7 |
Formula | C36H59N5O11 |
Molar Mass | 737.88 |
Transport | Room temperature in continental US; may vary elsewhere. |
Storage | Please store the product under the recommended conditions in the Certificate of Analysis. |
Reference | [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. [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. |