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Difference between revisions of "Glutamate"

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::1) Weigh 1.691 g L-Glutamic acid, monosodium salt hydrate, and transfer to 5 mL volumetric glass flask.
::1) Weigh 1.691 g L-Glutamic acid, monosodium salt hydrate, and transfer to 5 mL volumetric glass flask.
::2) Add 3 ml H<sub>2</sub>O (pH = 6).
::2) Add 3 ml H<sub>2</sub>O (pH = 6).  
::3) Neutralize with 5 N KOH.
::3) Check pH and adjust with 5N KOH to 7 if necessary.
::4) Check pH and adjust to 7 if necessary.
::4) Adjust final volume to 5 ml and divide into 0.5 ml portions.  
::5) Adjust final volume to 5 ml and divide into 0.5 ml portions.  
::5) Store frozen at -20 °C.
::6) Store frozen at -20 °C.





Revision as of 12:08, 17 December 2014


high-resolution terminology - matching measurements at high-resolution


Glutamate

Description

Glutamic acid

Glutamic acid, C5H9NO4, is an amino acid which occurs under physiological conditions mainly as the anion glutamate-, G, with pKa1 = 2.1, pKa2 = 4.07 and pKa3 = 9.47. Glutamate+malate is a CI-linked substrate combination, when glutamate is transported into the mt-matrix via the glutamate-aspartate carrier and reacts with oxaloacetate in the transaminase reaction to form aspartate and oxoglutarate. Glutamate as the sole substrate is transported by the electroneutral glutamate-/OH- exchanger, and is oxidized in the mitochondrial matrix by glutamate dehydrogenase to α-ketoglutarate ( 2-oxoglutarate). Ammonia (the byproduct of the reaction) passes freely through the mitochondrial membrane.

Abbreviation: G

Reference: Gnaiger 2012 MitoPathways



MitoPedia topics: Substrate and metabolite 

Application in HRR

G: Glutamate (L-Glutamic acid, monosodium salt hydrate, C5H8NO4Na; contains 1 mol H2O /mol); Sigma G 1626, 100 g, store at R.T.; FW = 169.1)

Preparation of 2 M stock solution (dissolved in H2O)

1) Weigh 1.691 g L-Glutamic acid, monosodium salt hydrate, and transfer to 5 mL volumetric glass flask.
2) Add 3 ml H2O (pH = 6).
3) Check pH and adjust with 5N KOH to 7 if necessary.
4) Adjust final volume to 5 ml and divide into 0.5 ml portions.
5) Store frozen at -20 °C.


Oxygraph-2k manual titrations MiPNet09.12 O2k-Titrations

  • Titration volume: 10 µl using a 25 µl syringe (2 ml O2k-chamber).
  • Final concentration: 10 mM.