Difference between revisions of "Antimycin A"
From Bioblast
m |
|||
Line 41: | Line 41: | ||
:::: '''Preparation of 10 mM stock solution''' (dissolved in 99.9% ethanol) | :::: '''Preparation of 10 mM stock solution''' (dissolved in 99.9% ethanol) | ||
::::# Weigh 10.8 mg of antimycin A and dissolve in 2 | ::::# Weigh 10.8 mg of antimycin A and dissolve in 2 mL of ethanol absolute (99.9%). | ||
::::# Divide into 0.2 mL portions in clear glass vials. | ::::# Divide into 0.2 mL portions in clear glass vials. | ||
::::# Store at -20 °C. | ::::# Store at -20 °C. |
Revision as of 15:18, 9 November 2020
Description
Antimycin A is an inhibitor of Complex III (CIII). It binds to the Qi site of CIII and inhibits the transfer of electrons from heme bH to oxidized Q (Qi site inhibitor). High concentrations of antimycin A also inhibit acyl-CoA oxidase and D-amino acid oxidase.
Abbreviation: Ama
Reference: MiPNet09.12
Application in HRR
- Ama: Antimycin A (from Streptomyces sp.) Sigma A 8674, 25 mg, store at -20 °C: FW = 540
- Caution: Very toxic. Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.
- Preparation of 5 mM stock solution (dissolved in 99.9% ethanol) for 2-mL O2k-chamber:
- Weigh 5.4 mg of antimycin A into a small glass vial. Caution: Ama is hygroscopic.
- Add 2 mL ethanol.
- Divide into 0.2 mL portions in clear glass vials.
- Store at -20 °C.
- » O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (2-mL O2k-chamber): 1 µL using a 10 µL Hamilton syringe.
- Final concentration: 2.5 µM.
- Preparation of 1,25 mM stock solution (dissolved in 99.9% ethanol) for 0.5-mL O2k-chamber:
- Take 100 µL of the 5 mM stock solution.
- Add 300 µL ethanol.
- » O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (0.5-mL O2k-chamber): 1 µL using a 10 µL Hamilton syringe.
- Final concentration: 2.5 µM.
- Microalgae applications
- For the specific case of microalgal cells it is recommended to prepare higher concentration:
- Preparation of 10 mM stock solution (dissolved in 99.9% ethanol)
- Weigh 10.8 mg of antimycin A and dissolve in 2 mL of ethanol absolute (99.9%).
- Divide into 0.2 mL portions in clear glass vials.
- Store at -20 °C.
- » O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume: 8 µL using a 10 µL syringe (2 mL O2k-Chamber).
- Final concentration: 40 µM.
Compare
- Myxothiazol - this CIII inhibitor induces less ROS production but is more expensive than Ama.
Antimycin A and ROS production
- Is hydrogen peroxide production measured after inhibition of the Electron transfer-pathway by rotenone and antimycin A of diagnostic significance? - Please, share your evidence (Email to [email protected]).
Inhibition of acyl-CoA oxidase and D-amino acid oxidase
- Vamecq J, Schepers L, Parmentier G, Mannaerts GP (1987) Inhibition of peroxisomal fatty acyl-CoA oxidase by antimycin A. Biochem J 248:603-7. - »Bioblast link«
MitoPedia topics:
Inhibitor