Difference between revisions of "TMPD"

Revision as of 16:55, 8 June 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 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

@@ Line 1: / Line 1: @@
 {{MitoPedia
 |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 [[Ascorbate|ascorbate]] and undergoes autooxidation as a function of [[oxygen pressure]], ascorbate and cytochrome c concentration.
+|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
 }}
@@ Line 9: / Line 13: @@
 |type=Substrate ETS
 }}
+== Application in [[HRR]]  ==
-== Application in [[HRR]]  ==
+=== Complex IV assay ===
+TMPD concentration: 0.5 mM.
-'''TMPD: N,N,N',N'-Tetramethyl-p-phenylenediamine dihydrochloride'''  (N,N,N',N'-Tetramethyl-p-phenylenediamine dihydrochloride,  C<sub>10</sub>H<sub>16</sub>N<sub>2</sub>*2HCl);  Sigma T 3134, 5 g, store at R.T.; FW = 237.2
+=== 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.
-'''Preparation of 200 mM stock solution''' (dissolved in H<sub>2</sub>O):
-::1) weigh 47.4 mg of N,N,N',N'-Tetramethyl-p-phenylenediamine dihydrochloride
+::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
-::3) add 0.8 M [[Ascorbate|ascorbate]] to a final concentration of 10 mM
+::3) Add 0.8 M [[Ascorbate|ascorbate]] to a final concentration of 10 mM
-::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