Seahorse - Mitochondrial Stress Test: Difference between revisions
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*** 1 mM Pyruvate (100 uL of a 1M stock) | *** 1 mM Pyruvate (100 uL of a 1M stock) | ||
** Re-adjust the pH to 7.4 (7.35-7.45) | ** Re-adjust the pH to 7.4 (7.35-7.45) | ||
* Wash cells twice with 500 | * Wash cells twice with 500 uL of pre-warmed media, add 500 uL final volume to each well | ||
* Incubate for 1h in non-CO2, humidified, 37C incubator to get rid of any extra CO2 | * Incubate for 1h in non-CO2, humidified, 37C incubator to get rid of any extra CO2 | ||
* While plate is incubating, prepare injection solutions in 1.8 mL of prepared XF Media. These are standard concentrations, and should be optimized for your system (especially FCCP) | * While plate is incubating, prepare injection solutions in 1.8 mL of prepared XF Media. These are standard concentrations, and should be optimized for your system (especially FCCP) | ||