ACSF Media
From Bridges Lab Protocols
Revision as of 12:39, 27 April 2017 by Davebridges (Talk | contribs) (Wrote initial table for buffers)
This media stock is the media for both the mitochondrial stress assays for skeletal muscle fibers. Because it is fairly well buffered, ECAR results are not reliable. Recheck the pH after warming and prior to each use. These volumes are to make 500 mL of media:
| Reagent | Stock Concentration | Volume (500 mL) | Final Concentration |
|---|---|---|---|
| Sodium chloride | 4 M | 15 mL | 120 mM |
| Potassium chloride | 1 M | 1.75 mL | 3.5 mM |
| Calcium Chloride | 1 M | 650 uL | 1.3 mM |
| Potassium chloride | 1 M | 2.7 mL | 5.4 mM |
| Potassium phosphate | 1M | 200 uL | 0.4 mM |
| Magnesium chloride | 1 M | 0.5 uL | 1 mM |
| HEPES | 1 M | 2.5 mL | 5 mM |
| D-Glucose | Powder | 1.35 g | 15 mM |
Add ~450 mL water, bring pH to 7.4. Bring final volume up to 500 mL and sterile filter into an autoclaved bottle. Store at 4C.
