Mitochondrial marker protein?
(by R.Jayakumar from roswellpark.org)
Thu Jun 21 13:24:07 EST 2007
It is a good idea to use VDAC as a loading control. VDAC is a outer
membrane pore protein and is generally present in stoichometric
concentrations on mitochondria.
From: methods-bounces from oat.bio.indiana.edu
[mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of Tom Anderson
Sent: Thursday, June 21, 2007 8:59 AM
To: methods from magpie.bio.indiana.edu
Subject: Re: Mitochondrial marker protein?
On Wed, 20 Jun 2007, WS wrote:
> any ideas for a good mitochondrial marker protein for human skeletal
> muscle which is easily to detect in westerns? a molecular weight above
> 25kd would be fine.
Prohibitin is a popular choice for this.
A colleague is working on the growth and replication of mitochondria,
and he's spent a lot of time thinking about how to quantify them. A
fundamental problem here is that the protein composition of mitochondria
changes according to signals and growth conditions, which means it's not
clear what 'amount' means - if you have the same amount of F0 ATPase but
twice as much of some matix enzyme, say SDH, do you have the same amount
of mitochondrion, or twice as much? Does it matter if some given mass of
proteins is divided between N mitochondria or 2N? I think he decided
that the gold standard was the number of copies of the mtDNA, but this
is a nightmare to measure; for most of his work, he's stained with
MitoTracker and measured the total volume of mitochondria in the cell.
I'll ask him if he has any suggestions about markers for blots, though.
You might also like to have a look at:
Tom Anderson, MRC Laboratory for Molecular Cell Biology, UCL, London
(t) +44 (20) 76797264 (f) +44 (20) 76797805 (e)
thomas.anderson from ucl.ac.uk
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