Succinate: Difference between revisions

Revision as of 11:08, 22 November 2010

high-resolution terminology - matching measurements at high-resolution

Succinate

Description

Succinate^2- 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

@@ Line 4: / Line 4: @@
 |info=[[MiPNet11.09]], [[MiPNet12.11]]
 |type=Respiration
-}}
+}} {{Labeling
-{{Labeling
 |discipline=Mitochondrial Physiology
 |enzymes=Complex II; Succinate Dehydrogenase
@@ Line 11: / Line 10: @@
 |instruments=Chemicals; Media
 |type=Respiration
 }}
-==HRR==
-S: Succinate disodium salt, hexahydrate, C4H4O4Na2.6H2O; FW=270.1 (Sigma, S 2378 (100 g) RT).
+== 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]]