Difference between revisions of "Oligomycin"
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|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. | |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. | ||
|info=[[MiPNet09.12]] | |info=[[MiPNet09.12]] | ||
}} | }} | ||
{{MitoPedia concepts | {{MitoPedia concepts | ||
|mitopedia concept=Respiratory state | |mitopedia concept=Respiratory state | ||
}} | }} | ||
{{MitoPedia topics | {{MitoPedia topics | ||
|mitopedia topic=Inhibitor | |mitopedia topic=Inhibitor | ||
}} | }} | ||
== Application in [[HRR]] == | == Application in [[HRR]] == | ||
'''Omy: Oligomycin''' (Oligomycin from ''Streptomyces diastatochromogenes''; mixture of oligomycins A, B, and C); Sigma O 4876, 5 mg; store at -20 °C; FW = 800. | :::: '''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. | :::: '''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): | ::: '''Preparation of 5 mM storage and stock solution''' (dissolved in 99.9% ethanol): | ||
:: | ::::# Weigh 4 mg oligomycin into a 2 ml glass vial. | ||
:: | ::::# Dissolve in 1 ml ethanol. | ||
:: | ::::# 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. | ||
:: | ::::# Store in glass vials (recommendation) at -20 °C. | ||
'''Oxygraph-2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | ::: '''Oxygraph-2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | ||
::* Titration volume: 1 µl using a 10 µl syringe (2 ml O2k-chamber). | ::::* Titration volume: 1 µl using a 10 µl syringe (2 ml O2k-chamber). | ||
::* Final concentration: 2 µg/ml (2.5 µM). | ::::* Final concentration: 2 µg/ml (2.5 µM). | ||
'''Problems with the use of oligomycin''' | ::: '''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 |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 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]]. | ||
[[Talk:Oligomycin|>>Discussion]] | [[Talk:Oligomycin|>>Discussion]] |
Revision as of 09:30, 16 May 2016
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 concepts:
Respiratory state
MitoPedia topics:
Inhibitor
Application in HRR
- 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):
- Weigh 4 mg oligomycin into a 2 ml glass vial.
- Dissolve in 1 ml ethanol.
- 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.
- Store in glass vials (recommendation) at -20 °C.
- Oxygraph-2k 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 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.
>>Discussion