From Bioblast
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
Application in HRR
- TMPD (C10H16N2*2HCl): Sigma T 3134, 5 g, store at RT; FW = 237.2.
- Caution: Light sensitive (store solution in a dark glass vial). Take care to minimize light and temperature exposure during use!
- Complex IV assay : For the Complex IV assay TMPD is required at 0.5 mM final concentration added from a 200 mM stock.
- TMPD (C10H16N2*2HCl): Sigma T 3134, 5 g, store at RT; FW = 237.2.
- Preparation of 200 mM stock solution (dissolved in H2O) for 2-mL O2k-chamber:
- Weigh 47.4 mg TMPD
- Dissolve in 1 mL H2O
- Divide into 0.2 mL portions
- Protect from light!
- Store frozen at -20 Β°C
- Preparation of 200 mM stock solution (dissolved in H2O) for 2-mL O2k-chamber:
- Β» O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (2-mL O2k-chamber): 5 Β΅L using a 25 Β΅L Hamilton syringe.
- Final concentration: 0.5 mM.
- Preparation of 250 mM stock solution (dissolved in H2O) for 0.5-mL O2k-chamber:
- Weigh 59,25 mg TMPD
- Dissolve in 1 mL H2O
- Divide into 0.2 mL portions
- Protect from light!
- Store frozen at -20 Β°C
- Β» O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (0.5-mL O2k-chamber): 1 Β΅L using a 10 Β΅L Hamilton syringe.
- Final concentration: 0.5 mM.
MitoPedia topics: Substrate and metabolite