From Bioblast
Description
Sodium vanadate (Na3VO4) is used as an ATPase inhibitor.
Abbreviation: VO4
Application in HRR
- VO4: orthovanadate (Sodium vanadate; Na3VO4), Sigma-Aldrich: S6508, store at RT, M = 183.91 gΒ·mol-1
- Preparation of 25 mM stock solution (dissolved in H2O)
- Weigh 45.98 mg.
- Under a fume hood, dissolve in a volume of distilled H2O lower than 10 mL, using a borosilicate Erlenmeyer flask or Falcon tube.
- Under the fume hood, adjust the pH of the solution to approximately 8.7 with HCl solution using pH test strips, the solution will turn yellow.
- Prepare a heated bath under the fume hood and allow the water to boil (e.g., with a heating plate and a borosilicate glass flask with water).
- Heat the solution in this bath until it turns colorless and allow it to cool to room temperature (it is possible to use an ice bath to make it faster).
- Readjust the pH to 8.7 with HCl under the fume hood, the solution will turn yellow again.
- Repeat this cycle of heating and readjusting the pH until the solution remains colorless at pH 8.7 and room temperature.
- Adjust the volume with distilled H2O to 10 mL.
- Divide in aliquots.
- Store at +4 Β°C.
- βThis treatment removes all decavanadate ions present in the Na3VO4 solution, which induces mitochondrial membrane depolarization and inhibition of oxygen consumptionβ, Chinopoulos 2014 Methods Enzymol
- Preparation of 25 mM stock solution (dissolved in H2O)
- Β» O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume: 2.4 Β΅L using a 10 Β΅L syringe (2 mL O2k-chamber).
- Final concentration: 30 Β΅M.
- Β» O2k manual titrations MiPNet09.12 O2k-Titrations
References
- [Chinopoulos 2014 Methods Enzymol]
MitoPedia topics: Inhibitor