TMPD: Difference between revisions
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{{MitoPedia | {{MitoPedia | ||
|abbr=Tm | |abbr=Tm | ||
|description='''N,N,N',N'-Tetramethyl-p-phenylenediamine dihydrochloride''' is applied as an artificial substrate for reducing [[cytochrome c]] in the respirometric assay for [[Complex IV|cytochrome c oxidase]] activity. It is maintained in a reduced state by [[ | |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 [[Complex IV|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. | ||
|info=MiPNet06.06, MiPNet09.12 | |info=[[MiPNet06.06]], [[MiPNet09.12]] | ||
|type=Substrate ETS | |||
}} | |||
{{MitoPedia methods | |||
|mitopedia method=Respirometry | |||
|type=Substrate ETS | |type=Substrate ETS | ||
}} | }} | ||
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|type=Substrate ETS | |type=Substrate ETS | ||
}} | }} | ||
== Application in [[HRR]] == | |||
== | === Complex IV assay === | ||
TMPD concentration: 0.5 mM. | |||
=== 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. | |||
::1) | ::1) Weigh 47.4 mg TMPD | ||
::2) | ::2) Dissolve in 1 ml H<sub>2</sub>O | ||
::3) | ::3) Add 0.8 M [[Ascorbate|ascorbate]] to a final concentration of 10 mM | ||
::4) | ::4) Divide into 0.2 ml portions | ||
::5) | ::5) Store frozen at -20 °C |
Revision as of 16:55, 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