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Suitable controls for for cellular fractionation


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#1 pobrien

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Posted 24 November 2009 - 06:57 AM

Hi everyone. Am new to this forum and was wondering could someone give me some advice.

I am performing an cellular fractionation experiment where I have isolated both nuclear and cytosolic fractions from Jurkat cells. I am pretty sure that the technique is working well but I need appropriate antibody controls looking for proteins found exclusively in the nucleus/cytosol (particularly the cytosol). I have used an anti-lamin B antibody to confirm that the nuclear fraction contains this protein but would like another control to be sure (plus i have run out of the antibody and have no more funds :angry: ).

Thanks in advance,

Phil

#2 bob1

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Posted 24 November 2009 - 03:49 PM

Nucleolin, histone H3 and tp53 are all found exclusively in the nucleus. Tubulin is found only in the cytoplasm.

#3 Janupagla

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Posted 05 December 2009 - 10:36 PM

I am performing an cellular fractionation experiment where I have isolated both well but I need appropriate antibody controls looking for proteins found. Cell fractionation by differential centrifugation and density equilibration on sucrose gradients ments. Corresponding positive availability of enzymes suitable as markers.


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