Difference between revisions of "Ascorbate"

Revision as of 11:25, 22 October 2020

high-resolution terminology - matching measurements at high-resolution

Ascorbate

Description

In respiratory assays for cytochrome c oxidase activity (Complex IV, CIV), ascorbate is added as regenerating system for maintaining TMPD in a reduced state. As has to be titrated into the respiration medium prior to the addition of TMPD, otherwise the reaction velocity of autoxidation is permanently elevated.

Abbreviation: As

Reference: MiPNet06.06

Application in HRR

As: Ascorbate sodium salt (Sodium L-ascorbate); Sigma A 7631, 100 g, store at RT; FW = 198.1.

Caution: Light sensitive (store solution in a dark glass vial).

Preparation of 800 mM stock solution (dissolved in H₂O) for 2-mL O2k-chamber:

Weigh 1.584 g of ascorbate sodium salt and dissolve in 10 mL H₂O
To prevent autoxidation, adjust pH to approx. 6 with ascorbic acid (a 137.6 mg.mL^-1 solution of pH 2).
Divide into 0.2 mL portions.
Store frozen at -20 °C.

»O2k manual titrations: MiPNet09.12

Titration volume (2-mL O2k-chamber): 5 µL using a 25 µL Hamilton syringe.
Final concentration: 2 mM.

Preparation of 1000 mM stock solution (dissolved in H₂O) for 0.5-mL O2k-chamber:

Weigh 1.980 g of ascorbate sodium salt and dissolve in 10 mL H₂O
To prevent autoxidation, adjust pH to approx. 6 with ascorbic acid (a 137.6 mg.mL^-1 solution of pH 2).
Divide into 0.2 mL portions.
Store frozen at -20 °C.

»O2k manual titrations: MiPNet09.12

Titration volume (0.5-mL O2k-chamber): 1 µL using a 10 µL Hamilton syringe.
Final concentration: 2 mM.

MitoPedia topics: Substrate and metabolite

@@ Line 12: / Line 12: @@
 :::: <span style="color:#8B008B"> '''Caution:''' Light sensitive (store solution in a dark glass vial).</span>
-::: '''Preparation of 800 mM stock solution''' (dissolved in H<sub>2</sub>O):
+<!--
+:::: '''Preparation of 800 mM stock solution''' (dissolved in H<sub>2</sub>O):
 ::::# Weigh 1.584 g of ascorbate sodium salt and dissolve in 10 mL H<sub>2</sub>O
 ::::# To prevent autoxidation, adjust pH to approx. 6 with ascorbic acid (a 137.6 mg.mL<sup>-1</sup> solution of pH 2).
 ::::# Divide into 0.2 mL portions.
 ::::# Store frozen at -20 °C.
 :::» '''O2k manual titrations:''' [[MiPNet09.12]]
@@ Line 24: / Line 24: @@
 ::::* Titration volume: 5 µL using a 25 µL syringe.
 ::::* Final conc. in 2 mL O2k-chamber: 2 mM.
+-->
+:::: '''Preparation of 800 mM stock solution''' (dissolved in H<sub>2</sub>O) for '''2-mL O2k-chamber''':
+::::# Weigh 1.584 g of ascorbate sodium salt and dissolve in 10 mL H<sub>2</sub>O
+::::# To prevent autoxidation, adjust pH to approx. 6 with ascorbic acid (a 137.6 mg.mL<sup>-1</sup> solution of pH 2).
+::::# Divide into 0.2 mL portions.
+::::# Store frozen at -20 °C.
+:::»'''O2k manual titrations:''' [[MiPNet09.12]]
+::::* Titration volume ('''2-mL O2k-chamber'''): 5 µL using a 25 µL Hamilton syringe.
+::::* Final concentration: 2 mM.
+:::: '''Preparation of 1000 mM stock solution''' (dissolved in H<sub>2</sub>O) for '''0.5-mL O2k-chamber''':
+::::# Weigh 1.980 g of ascorbate sodium salt and dissolve in 10 mL H<sub>2</sub>O
+::::# To prevent autoxidation, adjust pH to approx. 6 with ascorbic acid (a 137.6 mg.mL<sup>-1</sup> solution of pH 2).
+::::# Divide into 0.2 mL portions.
+::::# Store frozen at -20 °C.
+:::»'''O2k manual titrations:''' [[MiPNet09.12]]
+::::* Titration volume ('''0.5-mL O2k-chamber'''): 1 µL using a 10 µL Hamilton syringe.
+::::* Final concentration: 2 mM.
 {{MitoPedia topics
 |mitopedia topic=Substrate and metabolite
 }}