ACSF Media: Difference between revisions
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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" | ||
| Line 18: | Line 20: | ||
| 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 ]] | ||