Antimycin A: Difference between revisions

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:::: '''Preparation of 1,25 mM stock solution''' (dissolved in ethanol abs.) for '''0.5-mL O2k-chamber''':
:::: '''Preparation of 1.25 mM stock solution''' (dissolved in ethanol abs.) for '''0.5-mL O2k-chamber''':


::::# Take 100 ยตL of the 5 mM stock solution.
::::# Take 100 ยตL of the 5 mM stock solution.
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== Inhibition of acyl-CoA oxidase and D-amino acid oxidase ==
== 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. - [[Vamecq 1987 Biochem J |ยปBioblast linkยซ]]
::::* Vamecq J, Schepers L, Parmentier G, Mannaerts GP (1987) Inhibition of peroxisomal fatty acyl-CoA oxidase by antimycin A. Biochem J 248:603-7. - [[Vamecq 1987 Biochem J |ยปBioblast linkยซ]]
{{MitoPedia concepts}}
{{MitoPedia methods}}
{{MitoPedia O2k and high-resolution respirometry}}
{{MitoPedia topics
|mitopedia topic=Inhibitor
}}

Latest revision as of 13:23, 15 March 2022


high-resolution terminology - matching measurements at high-resolution


Antimycin A

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-Aldrich: A8674, store at -20 ยฐC, CAS: 1397-94-0, M = 540 gยทmol-1
Hazard statements: H300, H400; fatal if swallowed, very toxic to aquatic life
Sigma recommends to store desiccated, there is no hint to light sensitivity.
Light sensitivity of Antimycin A is reported by another vendor: AlfaAesar.


Preparation of 5 mM stock solution (dissolved in ethanol abs.) for 2-mL O2k-chamber:
  1. Weigh 5.4 mg of antimycin A into a small glass vial.
  2. Add 2 mL ethanol.
  3. Divide into 0.2 mL portions in clear glass vials.
  4. Store at -20 ยฐC.
ยป O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume (2-mL O2k-chamber): 1 ยตL using a 10 ยตL Hamilton syringe.
  • Final concentration: 2.5 ยตM.


Preparation of 1.25 mM stock solution (dissolved in ethanol abs.) for 0.5-mL O2k-chamber:
  1. Take 100 ยตL of the 5 mM stock solution.
  2. Add 300 ยตL ethanol.
ยป O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume (0.5-mL O2k-chamber): 1 ยตL using a 10 ยตL Hamilton syringe.
  • Final concentration: 2.5 ยตM.


Microalgae applications
For the specific case of microalgal cells it is recommended to prepare higher concentration:
Preparation of 10 mM stock solution (dissolved in ethanol abs.)
  1. Weigh 10.8 mg of antimycin A and dissolve in 2 mL of ethanol abs..
  2. Divide into 0.2 mL portions in clear glass vials.
  3. Store at -20 ยฐC.
ยป O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume (2-mL O2k-chamber): 8 ยตL using a 10 ยตL syringe.
  • Final concentration: 40 ยตM.


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. - ยปBioblast linkยซ







MitoPedia topics: Inhibitor 

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