Seahorse XF Media
This minimal media stock is the base media for both the glycolysis and mitochondrial stress assays. It contains no buffering and no carbon sources. It is a non-buffered media but still needs to be brought to pH 7.4 and checked before each assay.
For glycolysis assays add:
- 1-4 mM L-Glutamine
For mitochondrial assays add:
- 0.5-10 mM Sodium pyruvate
- 1-4 mM L-Glutamine
- 2-5 mM Glucose
To make base media add the following:
| Reagent | Stock Concentration | Volume (500 mL) | Final Concentration |
|---|---|---|---|
| Magnesium sulfate | 1 M | 400 uL | 0.8 mM |
| Calcium chloride | 1 M | 900 uL | 1.8 mM |
| Sodium Chloride | 4 M | 17.8 mL | 143 mM |
| Potassium chloride | 1 M | 2.7 mL | 5.4 mM |
| Sodium phosphate | 200 mM | 2.28 mL | 0.91 mM |
Add ~450 mL water, bring pH to 7.4 carefully (there is very little buffering capacity, so add NaOH slowly). Bring final volume up to 500 mL and sterile filter into an autoclaved bottle. Store at 4C.