Homogenization Buffer for Ca2+-Uptake Assay
| Reagent | Final concentration |
|---|---|
| K2HPO4 (pH 7.0) | 50 mm |
| NaF | 25 mm |
| Sucrose | 300 mm |
| EDTA (pH 7.0) | 1 mm |
| Dithiothreitol (DTT) | 0.5 mm |
Before use, add per 5 mL homogenization buffer:
-
5 µL 200 mm PMSF (in ethanol)
-
50 µL Protease inhibitor cocktail (in DMSO)
-
50 µL Phosphatase inhibitor cocktail (in DMSO)
The protease inhibitor cocktail should be diluted according to the manufacturer’s instructions. Stock solutions can be kept for several months when refrigerated, except DTT, PMSF, protease and phosphatase inhibitor cocktails, which should be kept at −20°C.
- © 2014 Cold Spring Harbor Laboratory Press
