Difference between revisions of "Malonate"

Revision as of 11:20, 25 November 2010

high-resolution terminology - matching measurements at high-resolution

Malonate

Description

Malonate (malonic acid) is a competitive inhibitor of Succinate dehydrogenase (Complex II).

Abbreviation: Mna

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.

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

Mna: Malonate (Malonic acid, CH₂(COOH)₂); Sigma M 1296, 5 g, 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 2 M stock solution (dissolved in H20): Recommended to prepare fresh every day

1) weigh 20.8 mg of Malonic acid directly into a Eppendorf tube of appropriate volume

2) dissolve in 20 µl H₂O

3) add 70 µl 5 N KOH and check pH

4) titrate small amounts (2 µl) of 5 N KOH until you reach a pH of 6.0 (approx. 4 titration steps). A pH of 7.0 would be ideal, however the pH jumps from pH 6.0 to pH 10.0 with only very little amounts of excess KOH.

5) add H₂O to a final volume of 100 µl

Oxygraph-2k Manual Titrations: MiPNet09.12

Titration volume: 5 µl using a 10 µl syringe
Final conc. in 2 ml O2k-chamber: 5 mM

@@ Line 23: / Line 23: @@
 ::1) weigh 20.8 mg of Malonic acid directly into a Eppendorf tube of appropriate volume
-::2) dissolve in 21 µl H20 + 79 µl 5 N KOH
+::2) dissolve in 20 µl H<sub>2</sub>O
-::3) check pH - should be around pH of 6.0. A pH of 7.0 would be ideal, however the pH jumps from pH 6.0 to pH 10.0 with only very little amounts of KOH (1 µl).
+::3) add 70 µl 5 N KOH and check pH
+::4) titrate small amounts (2 µl) of 5 N KOH until you reach a pH of 6.0 (approx. 4 titration steps). A pH of 7.0 would be ideal, however the pH jumps from pH 6.0 to pH 10.0 with only very little amounts of excess KOH.
+::5) add H<sub>2</sub>O to a final volume of 100 µl
 '''Oxygraph-2k Manual Titrations:'''  [[MiPNet09.12]]