Difference between revisions of "Ascorbate"
From Bioblast
Fontana Mona (talk | contribs) |
|||
| Line 17: | Line 17: | ||
::1) weigh 1.584 g of ascorbate sodium salt and dissolve in 10 ml H<sub>2</sub>O | ::1) weigh 1.584 g of ascorbate sodium salt and dissolve in 10 ml H<sub>2</sub>O | ||
::2) To prevent autoxidation, adjust pH to | ::2) To prevent autoxidation, adjust pH to approx. 6 with ascorbic acid (a 137.6 mg.ml<sup>-1</sup> solution of pH 2). | ||
::3) Divide into 0.2 ml portions. | ::3) Divide into 0.2 ml portions. | ||
::4) Store frozen at -20 °C. | ::4) Store frozen at -20 °C. | ||
Revision as of 11:19, 30 April 2013
- 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
MitoPedia topics: Substrate and metabolite
Application in HRR
As: Ascorbate sodium salt (Succinate disodium salt, hexahydrate, C6H706Na); Sigma A 4034, 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):
- 1) weigh 1.584 g of ascorbate sodium salt and dissolve in 10 ml H2O
- 2) To prevent autoxidation, adjust pH to approx. 6 with ascorbic acid (a 137.6 mg.ml-1 solution of pH 2).
- 3) Divide into 0.2 ml portions.
- 4) Store frozen at -20 °C.
Oxygraph-2k manual titrations: MiPNet09.12
- Titration volume: 5 µl using a 25 µl syringe
- Final conc. in 2 ml O2k-chamber: 2 mM
