Glutamate: Difference between revisions
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::::# 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 | ::::# Add 4 mL H<sub>2</sub>O. ย | ||
::::# | ::::# 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. ย | ::::# Adjust final volume to 5 mL and divide into 0.5 mL portions. ย | ||
::::# Store at -20 ยฐC. | ::::# Store at -20 ยฐC. | ||
:::ยป '''O2k manual titrations'''ย [[MiPNet09.12 O2k-Titrations]] | :::ยป '''O2k manual titrations'''ย [[MiPNet09.12 O2k-Titrations]] | ||
Revision as of 12:00, 5 November 2020
- high-resolution terminology - matching measurements at high-resolution
Glutamate
Description
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 ฮฑ-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
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)
- Weigh 1.691 g L-Glutamic acid, monosodium salt hydrate, and transfer to a 5 mL volumetric glass flask.
- Add 4 mL H2O.
- 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.
- Store at -20 ยฐC.
- ยป O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (2-mL O2k-chamber): 10 ยตL using a 25 ยตL Hamilton syringe.
- Titration volume (0.5-mL O2k-chamber): 2.5 ยตL using a 10 ยตL Hamilton syringe
- Final concentration: 10 mM.
MitoPedia topics: Substrate and metabolite
