NBD Ceramide
NBD Ceramide kdorfman Wed, 04/25/2018 - 14:32Invitrogen N22651
fluorescent marker for Golgi in live cells
Ex: 466nm Em: 536nm
Follow instructions attached: Add 150 µL sterile H2O to the 5 mg in the bottle. (Makes 0.5mM (=500 µM) in BSA)
Aliquot 10 µL, and freeze
Working concentration is 5 µM
Add 990 µL HBSS (Hank's)(0.34 mg/mL BSA) to 10 µL in tube
Makes enough for 8 groups.
Aliquot ~120 µL per group, enough to cover the small circular coverslip in the viewing dish.
To use:
- Rinse with HBSS (see notes)
- Treat with ceramide solution
- incubate in cold 30 min
- rinse 3x in cold HBSS
- replace HBSS with non-CO2 medium, incubate ~20 minutes
NOTES:
Minimal rinse! Very gently! Cold cells will fall off!
If following with ER tracker, rinse with HBSS, then replace with warm non-CO2 medium
If following with Mitotracker, rinse with PBS, then replace with warm non-CO2 medium