Nuclear permeability during apoptosis

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hima12
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Nuclear permeability during apoptosis

Hello,
Is there any kit available to determine the nuclear permeability potential changes during apoptosis similar to the mitochondrial membrane permeability potential changes?
Thanks in advance.
Hima.
 

The FFM
The FFM's picture
I couldnt find a kit, but

I couldnt find a kit, but here are papers with protocols for the assay to do it all yourself
 

Anal Biochem. 2003 Jul 15;318(2):244-53.

In vitro measurement of nuclear permeability changes in apoptosis.

Roehrig S, Tabbert A, Ferrando-May E.

Molecular Toxicology Group, Department of Biology, University of Konstanz, P.O.
Box X911, 78457 Konstanz, Germany.

In the eukaryotic cell, exchange of biomolecules between nucleus and cytoplasm is
a highly regulated process which responds sensitively to changes of the
environment. One well-known cellular response to environmental challenges is cell
death by apoptosis. In fact, apoptosis has been shown to affect the
nucleocytoplasmic transport machinery, in particular the nuclear pore, by
modulating its size exclusion limit for passive diffusion. The underlying
molecular factors are still unknown, mainly because of the lack of a suitable
system to detect and quantitate the apoptotic effects on the nuclear pore. Here
we present an assay that was designed to measure alterations of the permeability
of the nuclear envelope under apoptotic conditions. The assay is based on the
well-established technique of selective permeabilization of the plasma membrane
with digitonin and allows assessment of permeability changes in nonfixed samples.
It comprises a computer program, called Nuclear Permeability Assay, for the
quantitation of the nuclear fluorescence signal, which may be generally employed
for the evaluation of in vitro transport systems using semipermeabilized cells,
such as assays for nuclear import and export.

PMID: 12814628

You can Purchase the Digitonin from Sigma-Aldrich
Also
In vitro assay for the quantitation of apoptosis-induced alterations of nuclearenvelope permeability. [Nat Protoc. 2006] PMID:17406565

Caspases mediate nucleoporin cleavage, but not early redistribution of nucleartransport factors and modulation of nuclear permeability in apoptosis. [CellDeath Differ. 2001] PMID:11423910

hima12
hima12's picture
Thanks FFM

Thanks FFM
I appreciate your answer.

heehawmcduff
heehawmcduff's picture
The paper that FFM described

The paper that FFM described is probably your best source but this may also help too. 
http://jvi.asm.org/cgi/content/abstract/83/7/3150
I know that people have also lightly permeablised cells and looked at the influx of texas-red labelled dextran particles from the cytoplasm through the nuclear membrane as a result of treatment.

hima12
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Thank you Heehawmcduff.

Thank you Heehawmcduff.