TMPD: Difference between revisions
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|info=[[MiPNet06.06]], [[MiPNet09.12]] | |info=[[MiPNet06.06]], [[MiPNet09.12]] | ||
|type=Substrate ETS | |type=Substrate ETS | ||
}} | }}{{MitoPedia methods | ||
{{MitoPedia methods | |||
|mitopedia method=Respirometry | |mitopedia method=Respirometry | ||
|type=Substrate ETS | |type=Substrate ETS | ||
}} | }}{{MitoPedia topics | ||
{{MitoPedia topics | |||
|mitopedia topic=Substrate and metabolite | |mitopedia topic=Substrate and metabolite | ||
|type=Substrate ETS | |type=Substrate ETS | ||
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=== Preparation of 200 mM stock === | === Preparation of 200 mM stock === | ||
TMPD (C<sub>10</sub>H<sub>16</sub>N<sub>2</sub>*2HCl): Sigma T 3134, 5 g, store at RT; FW = 237.2. | TMPD (C<sub>10</sub>H<sub>16</sub>N<sub>2</sub>*2HCl): Sigma T 3134, 5 g, store at RT; FW = 237.2. | ||
::1) Weigh 47.4 mg TMPD | ::1) Weigh 47.4 mg TMPD | ||
::2) Dissolve in 1 ml H<sub>2</sub>O | ::2) Dissolve in 1 ml H<sub>2</sub>O |
Revision as of 17:23, 8 June 2012
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 activity. It is maintained in a reduced state by ascorbate and undergoes autoxidation as a function of oxygen pressure, 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
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