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Difference between revisions of "Glutamate"

From Bioblast
(no KOH is necessary when using the monosodium salt hydrate)
Line 15: Line 15:


::::# Weigh 1.691 g L-Glutamic acid, monosodium salt hydrate, and transfer to a 5 mL volumetric glass flask.
::::# Weigh 1.691 g L-Glutamic acid, monosodium salt hydrate, and transfer to a 5 mL volumetric glass flask.
::::# Add 3 mL H<sub>2</sub>O.  
::::# Add 4 mL H<sub>2</sub>O.  
::::# Adjust pH with 5 M KOH to pH 7.0.
::::# Check pH. Usually it is ph 7.0. If not, adjust with 5 M KOH to pH 7.0.
::::# Adjust final volume to 5 mL and divide into 0.5 mL portions.  
::::# Adjust final volume to 5 mL and divide into 0.5 mL portions.  
::::# Store at -20 °C.
::::# Store at -20 °C.


:::» '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
:::» '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]

Revision as of 11:00, 5 November 2020


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 substrate combination supporting an N-linked pathway control state, 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), representing the glutamate-anaplerotic pathway control state. Ammonia (the byproduct of the reaction) passes freely through the mitochondrial membrane.

Abbreviation: G

Reference: Gnaiger 2014 MitoPathways

G

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 a 5 mL volumetric glass flask.
  2. Add 4 mL H2O.
  3. Check pH. Usually it is ph 7.0. If not, adjust with 5 M KOH to pH 7.0.
  4. Adjust final volume to 5 mL and divide into 0.5 mL portions.
  5. Store at -20 °C.
» O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume (2-mL O2k-chamber): 10 µL using a 25 µL Hamilton syringe.
  • Titration volume (0.5-mL O2k-chamber): 2.5 µL using a 10 µL Hamilton syringe
  • Final concentration: 10 mM.

MitoPedia topics: Substrate and metabolite