Difference between revisions of "Talk:TMPD"
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=== Complex IV assay === | === Complex IV assay === | ||
* [[Complex IV]] | * For the [[Complex_IV#HRR_protocol:_Complex_IV_assay|Complex IV assay]] TMPD is required at 0.5 mM final concentration added from a 200 mM stock. | ||
=== Preparation of 200 mM stock === | === Preparation of 200 mM stock === | ||
Revision as of 10:05, 20 March 2014
Complex IV assay
- For the Complex IV assay TMPD is required at 0.5 mM final concentration added from a 200 mM stock.
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) Divide into 0.2 ml portions
- 4) Protect from light!
- 5) Store frozen at -20 °C
Caution: Light sensitive (store solution in a dark glass vial). Take care to minimize light and temperature exposure during use!
It is important to add Ascorbate before TMPD to avoid uncontrollable autoxidation. Ascorbate maintains TMPD in a reduced state. Electrons flow from TMPD to cytochrome c.
For preparation and titration please visit Ascorbate
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
