I'm not sure either on the mechanism of Mitotracker Red CMXRos, I've utilised it to stain mouse ovarian follicles.
-- What I did was to fix the follicles then stain them.. I found that the stain was still retained in the fixed cells of the follicle. Not sure what to make of all of that.. Everyone is saying it is metabolised and retained. My staining protocol involved several washing steps aswell as counter staining with DAPI and both dyes were successfully retained, DAPI was expected to be but after reading all of this I would have expected Mitotracker to be washed out considering the follicles were fixed..
Hmm... Guess it is retained when staining post-fixation as well?
Anyone know what the deal is?
On Saturday, October 24, 2009 4:22:17 AM UTC+10:30, Nathan McCorkle wrote:
On Saturday, October 24, 2009 4:22:17 AM UTC+10:30, Nathan McCorkle wrote:
Take a look at these, I think once it gets into the mito it gets
metabolized and then it stays there... Not sure, I can look more at
the last link in this list in a few hours, but gotta go to a thesis
defense on 3d protein visualisation now
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi= B6WBK-4P6M8P4-4&_user=47004&_ rdoc=1&_fmt=&_orig=search&_ sort=d&_docanchor=&view=c&_ searchStrId=1061331114&_ rerunOrigin=google&_acct= C000005018&_version=1&_ urlVersion=0&_userid=47004& md5= 4a0d94df2e60a2bc218dd3cde5180f 17
http://molpharm.aspetjournals.org/cgi/content/abstract/45/6/ 1145
http://bbc.mcw.edu/Computation/models/text/ Mitochondrial%20Inner% 20Membrane% 20Electrophysiology% 20Assessed%20by%20Rhodamine- 123%20Transport%20and% 20Fluorescence.pdf
On Fri, Oct 23, 2009 at 11:26 AM, J. S. John <phill...@gmail.com> wrote:
>
> On Fri, Oct 23, 2009 at 12:11 AM, Nathan McCorkle <nmz...@gmail.com> wrote:
>
>> No I think they are just intra-mito proteins, not necessarily membrane
>> bound, but potentially, the data sheet says the dye gets sequestered
>> into the mito, so it could then interact with any protein inside,
>> including membrane ones.
>
> What makes Mito tracker specific to the mitochondria membrane?
> Wouldn't it bind to the cell membrane proteins also?
>
> >
>
--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics
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