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Actin expression in protein extracted from tissue

Actin expression in protein extracted from tissue - Protocols and Methods Forum

Actin expression in protein extracted from tissue - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 04-02-2009, 10:31 PM
Peter Ellis
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Default Actin expression in protein extracted from tissue



DK wrote:

RNALater is largely an extremely concentrated ammonium sulphate
solution. It preserves RNA by precipitating out the RNAses in the
tissue. Without knowing the composition of your lysis buffer, it's hard
to say whether you would have re-solubilised the actin in the sample.

Peter
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Old 04-02-2009, 11:05 PM
DK
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Default Actin expression in protein extracted from tissue

In article <[Only registered users see links. ].net> , "Kamalian, Laleh" <[Only registered users see links. ].uk> wrote:

Since you are using a solution for which composition is unknown,
it is impossible to say what might have happened to the cells/proteins
in your samples during the storage...


Since you are using a solution for which composition is unknown,
it is impossible to say whether it was an appropriate solution for
actin extraction.

Sonication of small volumes is difficult to control. Some samples
might have been fried while others not completely extracted.



One of the centrifuges I have delivers 12,000 g at full speed and
another 400,000 g. What about yours? That might make a lot of
difference for actin. Actin's solubility depends on salt concentration.
Do you know how much and what kinds of salt was present in your
solution?


I'd say preserve your samples properly and don't rely on magic
black boxes provided by companies. Then your results can be interpreted
by you and others.

DK
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