NBD Ceramide

Invitrogen 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