mitochondria concentration - (May/06/2008 )

Hello there,

Can anyone provide me some directions on how to determine the concentration of mitochondria?

I need to know how much mitochondria i have for my protease digest and western blot assays.

Thank you

Cheers

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We just use BCA kit to determine protein content.

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In that case, can i just use a simple Bradford assay?

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instead of protein, you may measure relative physiological activity by MTT or MTS

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Is there a way to correlate the relative physiological activity to the concentration of mitochondria?

Thanks!

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so you need to calibrate your system; I suggest to count a suspension of mitochondria and correlate with their physiological activity or protein concentration

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Thanks all!

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There are some dyes which bind to mitochondria (MitoSOX-Red? Invitrogen) depending on the membrane potential. Flow cytometry or immunofluorescence analysis will help in estimating mitochondrial density.

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Some assays here,
http://search.vadlo.com/b/q?sn=158621799&a...assay&rel=0

but I think measuring the protein concentration of mitochondrial fraction should be sufficient.


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