Difference between revisions of "TMPD"
From Bioblast
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::4) Divide into 0.2 ml portions | ::4) Divide into 0.2 ml portions | ||
::5) Store frozen at -20 °C | ::5) 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: 0.5 mM | |||
Revision as of 17:17, 21 November 2012
- high-resolution terminology - matching measurements at high-resolution
TMPD
Description
N,N,N',N'-Tetramethyl-p-phenylenediamine dihydrochloride, TMPD, is applied as an artificial substrate for reducing cytochrome c in the respirometric assay for cytochrome c oxidase (CIV) activity. It is maintained in a reduced state by ascorbate and undergoes autoxidation as a function of oxygen pressure, TMPD, ascorbate and cytochrome c concentration.
Abbreviation: Tm
Reference: MiPNet06.06, MiPNet09.12
MitoPedia methods:
Respirometry
MitoPedia topics:
Substrate and metabolite
Application in HRR
Complex IV assay
- Complex IV - TMPD concentration: 0.5 mM.
Preparation of 200 mM stock
TMPD (C10H16N2*2HCl): Sigma T 3134, 5 g, store at RT; FW = 237.2.
- 1) Weigh 47.4 mg TMPD
- 2) Dissolve in 1 ml H2O
- 3) Add 0.8 M ascorbate to a final concentration of 10 mM
- 4) Divide into 0.2 ml portions
- 5) 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: 0.5 mM
