Difference between revisions of "Succinate"
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|info=[[MiPNet11.09]], [[MiPNet12.11]] | |info=[[MiPNet11.09]], [[MiPNet12.11]] | ||
|type=Respiration | |type=Respiration | ||
}} | }} {{Labeling | ||
{{Labeling | |||
|discipline=Mitochondrial Physiology | |discipline=Mitochondrial Physiology | ||
|enzymes=Complex II; Succinate Dehydrogenase | |enzymes=Complex II; Succinate Dehydrogenase | ||
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|instruments=Chemicals; Media | |instruments=Chemicals; Media | ||
|type=Respiration | |type=Respiration | ||
}} | }} | ||
== Application in [[HRR]] == | |||
Stock solution: 1 000 mM (1 M; 2.701 g/10 ml H2O); Weigh 2.701 g of succinate disodium salt, add H2O to final volume 10 ml, check pH with pH paper and adjust to 7.0 if necessary. Divide into 0.5 ml portions. Store frozen at -20 °C. [[MiPNet03.02]] | '''S: Succinate''' (Succinate disodium salt, hexahydrate, C<sub>4</sub>H<sub>6</sub>0<sub>5</sub>; FW=270.1 (Sigma, S 2378 (100 g) RT). | ||
, C<sub>4</sub>H<sub>6</sub>0<sub>5</sub>; Sigma M 1000(R.T.): FW = 134.1) | |||
'''Preparation of 500 mM L-Malic acid solution''' (dissolved in H<sub>2</sub>O): | |||
::1) weigh 537 mg of L-Malic acid and transfer to 5 mL volumetric glass flask | |||
::2) add 3 ml H<sub>2</sub>O | |||
::3) neutralize with 10 N KOH (approx. 750 µl) | |||
::4) check pH and adjust to 7 if necessary (add approx. another 2 µl 10 N KOH) | |||
::5) adjust final volume to 5 ml and divide into 0.2 ml portions. | |||
::6) store frozen at -20 °C | |||
'''Oxygraph-2k Manual Titrations:''' [[MiPNet09.12]] | |||
::* Titration volume: 5 µl using a 25 µl syringe | |||
::* Final conc. in 2 ml O2k-chamber: 2 mM | |||
== HRR == | |||
S: Succinate disodium salt, hexahydrate, C4H4O4Na2.6H2O; FW=270.1 (Sigma, S 2378 (100 g) RT). | |||
Stock solution: 1 000 mM (1 M; 2.701 g/10 ml H2O); Weigh 2.701 g of succinate disodium salt, add H2O to final volume 10 ml, check pH with pH paper and adjust to 7.0 if necessary. Divide into 0.5 ml portions. Store frozen at -20 °C. [[MiPNet03.02]] | |||
Final conc. in 2 ml O2k-chamber: 10 mM; titrate 20 µl using the 50 µl syringe. [[MiPNet09.12]] | Final conc. in 2 ml O2k-chamber: 10 mM; titrate 20 µl using the 50 µl syringe. [[MiPNet09.12]] |
Revision as of 11:08, 22 November 2010
Description
Succinate2- is formed in the TCA cycle, and is a substrate of CII, reacting to fumarate and feeding electrons into the Q-junction through flavin adenine dinucleotide (FADH2). Succinate supports electron flux exclusively through Complex II via FADH2.
Abbreviation: S
Reference: MiPNet11.09, MiPNet12.11
Labels:
Enzyme: Complex II; Succinate Dehydrogenase"Complex II; Succinate Dehydrogenase" is not in the list (Adenine nucleotide translocase, Complex I, Complex II;succinate dehydrogenase, Complex III, Complex IV;cytochrome c oxidase, Complex V;ATP synthase, Inner mt-membrane transporter, Marker enzyme, Supercomplex, TCA cycle and matrix dehydrogenases, ...) of allowed values for the "Enzyme" property. Regulation: Substrate; Glucose; TCA Cycle"Substrate; Glucose; TCA Cycle" 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
S: Succinate (Succinate disodium salt, hexahydrate, C4H605; FW=270.1 (Sigma, S 2378 (100 g) RT).
, C4H605; Sigma M 1000(R.T.): FW = 134.1)
Preparation of 500 mM L-Malic acid solution (dissolved in H2O):
- 1) weigh 537 mg of L-Malic acid and transfer to 5 mL volumetric glass flask
- 2) add 3 ml H2O
- 3) neutralize with 10 N KOH (approx. 750 µl)
- 4) check pH and adjust to 7 if necessary (add approx. another 2 µl 10 N KOH)
- 5) adjust final volume to 5 ml and divide into 0.2 ml portions.
- 6) store frozen at -20 °C
Oxygraph-2k Manual Titrations: MiPNet09.12
- Titration volume: 5 µl using a 25 µl syringe
- Final conc. in 2 ml O2k-chamber: 2 mM
HRR
S: Succinate disodium salt, hexahydrate, C4H4O4Na2.6H2O; FW=270.1 (Sigma, S 2378 (100 g) RT).
Stock solution: 1 000 mM (1 M; 2.701 g/10 ml H2O); Weigh 2.701 g of succinate disodium salt, add H2O to final volume 10 ml, check pH with pH paper and adjust to 7.0 if necessary. Divide into 0.5 ml portions. Store frozen at -20 °C. MiPNet03.02
Final conc. in 2 ml O2k-chamber: 10 mM; titrate 20 µl using the 50 µl syringe. MiPNet09.12