350 ml 0.1 M PB solution (pH 7.35)
150 ml ethylene glycol
100 μg sodium azide
150 g sucrose
Final cryoprotectant concentration will be: 30% ethylene glycol, 30% sucrose, 0.00002% sodium azide, in 0.1 M PB (pH 7.35). Prepare this solution under the hood.
- Pour 250 ml of 0.1 M PB into 1 L flask.
- Pour 150 ml of ethylene glycol into the flask.
- Add 100 μg sodium azide.
- Stir for 30 min
- Add 150 g of sucrose
- Stir for 2 hr
- Bring volume up to 500 ml with 0.1 M PB
- Store cryoprotectant solution in refrigerator (+4ºC)