Na+-Free Extracellular Saline Containing Ionophores
| Reagent | Amount to add | Final concentration |
|---|---|---|
| KCl | 134.0 mg | 182.5 mm |
| HEPES buffer | 29.8 mg | 12.5 mm |
| Mg-ATP | 25.4 mg | 5 mm |
| Na-GTP | 2.6 mg | 0.5 mm |
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1. Combine the above ingredients in 8 mL of ultrapure water. Adjust the pH to 7.3 with KOH. The osmolarity should be 365 mOsmol/L. Adjust to a final volume of 10 mL using ultrapure water. Aliquot into 1-mL portions, and store at –20°C for up to 1 wk.
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2. Just before use, add the following ionophores to the extracellular saline solution: 3 µm gramicidin, 10 µm monensin, and 100 µm ouabain.
- © 2012 Cold Spring Harbor Laboratory Press
