Difference between revisions of "ACSF Media"
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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. | 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: | + | |
| + | This recipe and protocol for isolating/measuring muscle fibers is found [http://www.agilent.com/cs/pubimages/misc/TB_Intact_Muscle_Fibers.pdf here] | ||
| + | Recheck the pH after warming and prior to each use. These volumes are to make '''500 mL''' of media: | ||
{| class="wikitable" | {| class="wikitable" | ||
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| 3.5 mM | | 3.5 mM | ||
|- | |- | ||
| − | | Calcium | + | | Calcium chloride |
| 1 M | | 1 M | ||
| 650 uL | | 650 uL | ||
| 1.3 mM | | 1.3 mM | ||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
|- | |- | ||
| Potassium phosphate | | Potassium phosphate | ||
| − | | | + | | 0.2 M |
| − | | | + | | 1 mL |
| 0.4 mM | | 0.4 mM | ||
|- | |- | ||
| Line 51: | Line 48: | ||
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. | 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. | ||
| − | [[ Category: | + | [[ Category: Buffer ]] |
[[ Category: Seahorse ]] | [[ Category: Seahorse ]] | ||
[[ Category: Cell Culture ]] | [[ Category: Cell Culture ]] | ||
Latest revision as of 14:28, 2 August 2017
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.
This recipe and protocol for isolating/measuring muscle fibers is found here 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 phosphate | 0.2 M | 1 mL | 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.
