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Difference between revisions of "Oligomycin"

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::::* Several researchers have reported problems associated with the use of oligomycin which appear primarily related to difficulties with getting rid of the inhibitor after application in a respirometry chamber. In our hands following rigouros washing procedures generally helps to avoid such problems: for more details, see [[MiPNet19.03_O2k-cleaning_and_ISS |MiPNet19.03_O2k-cleaning_and_ISS]]. Solutions suggested by our users in the [[O2k-Network_discussion_forum|O2k-Network Discussion forum]] can be found on the [[Talk:Oligomycin|Discussion  page]]
::::* Several researchers have reported problems associated with the use of oligomycin which appear primarily related to difficulties with getting rid of the inhibitor after application in a respirometry chamber. In our hands following rigouros washing procedures generally helps to avoid such problems: for more details, see [[MiPNet19.03_O2k-cleaning_and_ISS |MiPNet19.03_O2k-cleaning_and_ISS]]. Solutions suggested by our users in the [[O2k-Network_discussion_forum|O2k-Network Discussion forum]] can be found on the [[Talk:Oligomycin|Discussion  page]]


::::* Oligomycin has frequently been shown to lower subsequently measured ETS capacity, a problem also reported in the [[Talk:Oligomycin#ETS_capacity_after_inhibition_by_Omy_.282013-02-21.29|O2k-Network Discussion forum]]. A recent paper by [[Ruas_2016_PLOS_ONE|Ruas et al. (2016)]] adressed this issue systematically and reports detailed results also on (unfortunately non-successfull) suggested alternatives to oligomycin. A potentially simple solution for at least some cell types has been suggested by [[Garcia_e_Souza_Luiz_Felipe |Luiz Felipe Garcia e Souza]], see [[Talk:Oligomycin#ETS_capacity_after_inhibition_by_Omy_.282013-02-21.29|here]].
::::* Oligomycin has frequently been shown to lower subsequently measured ET capacity, a problem also reported in the [[http://wiki.oroboros.at/index.php/Talk:Oligomycin#ET_capacity_after_inhibition_by_Omy_.282013-02-21.29|O2k-Network Discussion forum]]. A recent paper by [[Ruas_2016_PLOS_ONE|Ruas et al. (2016)]] adressed this issue systematically and reports detailed results also on (unfortunately non-successfull) suggested alternatives to oligomycin. A potentially simple solution for at least some cell types has been suggested by [[Garcia_e_Souza_Luiz_Felipe |Luiz Felipe Garcia e Souza]], see [[http://wiki.oroboros.at/index.php/Talk:Oligomycin#ET_capacity_after_inhibition_by_Omy_.282013-02-21.29|here]].
::::* Oligomycin has been found ineffective as inhibitor of the  ATP synthase in various strains of yeast, a problem we were made aware of by [[Agrimi_G |Gennaro Agrimi]] and which is briefly described and discussed [[Talk:Oligomycin#Lack_of_inhibition_with_Omy_in_yeast_cells_.282014-08-07.29| here]].
::::* Oligomycin has been found ineffective as inhibitor of the  ATP synthase in various strains of yeast, a problem we were made aware of by [[Agrimi_G |Gennaro Agrimi]] and which is briefly described and discussed [[Talk:Oligomycin#Lack_of_inhibition_with_Omy_in_yeast_cells_.282014-08-07.29| here]].



Revision as of 08:36, 20 October 2017


high-resolution terminology - matching measurements at high-resolution


Oligomycin

Description

Oligomycin (Omy) is an inhibitor of ATP synthase by blocking its proton channel (Fo subunit), which is necessary for oxidative phosphorylation of ADP to ATP (energy production). The inhibition of ATP synthesis also inhibits respiration. In OXPHOS analysis, Omy is used to induce a LEAK state of respiration.

Abbreviation: Omy

Reference: MiPNet09.12


MitoPedia topics: Inhibitor 

Application in HRFR

Omy: Oligomycin (Oligomycin from Streptomyces diastatochromogenes; mixture of oligomycins A, B, and C); Sigma O 4876, 5 mg; store at -20 °C; FW = 800.
Caution: Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.


Preparation of 5 mM storage and stock solution (dissolved in 99.9% ethanol):
  1. Weigh 4 mg oligomycin into a 2 ml glass vial.
  2. Dissolve in 1 ml ethanol.
  3. Divide into 0.2 ml portions or store in 2 ml glass vial and use subsamples during experiments to avoid contamination of the storage solution.
  4. Store in glass vials (recommendation) at -20 °C.


O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume: 1 µl using a 10 µl syringe (2 ml O2k-chamber).
  • Final concentration: 2 µg/ml (2.5 µM).


Problems with the use of oligomycin
  • Several researchers have reported problems associated with the use of oligomycin which appear primarily related to difficulties with getting rid of the inhibitor after application in a respirometry chamber. In our hands following rigouros washing procedures generally helps to avoid such problems: for more details, see MiPNet19.03_O2k-cleaning_and_ISS. Solutions suggested by our users in the O2k-Network Discussion forum can be found on the Discussion page
  • Oligomycin has frequently been shown to lower subsequently measured ET capacity, a problem also reported in the [Discussion forum]. A recent paper by Ruas et al. (2016) adressed this issue systematically and reports detailed results also on (unfortunately non-successfull) suggested alternatives to oligomycin. A potentially simple solution for at least some cell types has been suggested by Luiz Felipe Garcia e Souza, see [[1]].
  • Oligomycin has been found ineffective as inhibitor of the ATP synthase in various strains of yeast, a problem we were made aware of by Gennaro Agrimi and which is briefly described and discussed here.


References

  • Estabrook_1961_Biochem Biophys Res Commun


>>Discussion