Difference between revisions of "Antimycin A"
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== Application in [[HRR]] == | |||
'''Ama: Antimycin A''' (Antimycin A from ''Streptomyces'' sp., Sigma A 8674, 25 g, store at -20 °C): FW = 540) | |||
<span style="color:#8B008B"> '''Caution:''' Very toxic. Chemicals stored in the fridge or freezer should be adjusted to room temperature before opening.</span> | |||
'''Preparation of 5 mM stock solution''' (dissolved in 99.9% Ethanol): | |||
::1) weigh 5.5 mg of Antimycin A directly in to 2 mL glass vial !Difficult to weigh - appears to be hydroscopic! | |||
::2) add 2 ml Ethanol | |||
::3) divide into 0.2 ml portions | |||
::4) store in glass vial (recommendation) at -20 °C | |||
!White precipitate will occur, which will be dissolved after 1-2 min of stirring on magnetic stirrer at room temperature! | |||
::3) neutralize with 5 N KOH (approx. 465 µl) !ADP will dissolve after addition of KOH! (this is '''Solution A''') | |||
::4) check pH and adjust to 7 if necessary | |||
::5) adjust final volume to 2 ml and divide into 0.2 ml portions | |||
::6) store frozen at -80 °C | |||
'''Oxygraph-2k Manual Titrations:''' [[MiPNet09.12]] | |||
::* Titration volume: 8-20 µl using a 25 µl syringe | |||
::* Final conc. in 2 ml O2k-chamber: 2-5 mM | |||
Revision as of 12:40, 22 November 2010
- high-resolution terminology - matching measurements at high-resolution
Antimycin A
Description
Inhibitor of Complex III.
Abbreviation: Ama
Reference: MiPNet09.12,
Labels:
Regulation: Respiration; OXPHOS; ETS Capacity"Respiration; OXPHOS; ETS Capacity" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property., Flux Control; Additivity; Threshold; Excess Capacity"Flux Control; Additivity; Threshold; Excess Capacity" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property.
HRR: Chemicals; Media"Chemicals; Media" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property.
Application in HRR
Ama: Antimycin A (Antimycin A from Streptomyces sp., Sigma A 8674, 25 g, store at -20 °C): FW = 540)
Caution: Very toxic. Chemicals stored in the fridge or freezer should be adjusted to room temperature before opening.
Preparation of 5 mM stock solution (dissolved in 99.9% Ethanol):
- 1) weigh 5.5 mg of Antimycin A directly in to 2 mL glass vial !Difficult to weigh - appears to be hydroscopic!
- 2) add 2 ml Ethanol
- 3) divide into 0.2 ml portions
- 4) store in glass vial (recommendation) at -20 °C
!White precipitate will occur, which will be dissolved after 1-2 min of stirring on magnetic stirrer at room temperature!
- 3) neutralize with 5 N KOH (approx. 465 µl) !ADP will dissolve after addition of KOH! (this is Solution A)
- 4) check pH and adjust to 7 if necessary
- 5) adjust final volume to 2 ml and divide into 0.2 ml portions
- 6) store frozen at -80 °C
Oxygraph-2k Manual Titrations: MiPNet09.12
- Titration volume: 8-20 µl using a 25 µl syringe
- Final conc. in 2 ml O2k-chamber: 2-5 mM
