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Detergent concentration for nuclear membrane dissolving

Detergent concentration for nuclear membrane dissolving - Protocols and Methods Forum

Detergent concentration for nuclear membrane dissolving - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 02-15-2009, 08:29 PM
Werner Straube
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Default Detergent concentration for nuclear membrane dissolving



Hello everybody,

A friend of mine is interested isolating nuclear proteins. After he has
isolated the nuclei of cells he then wants to dissolve the nuclear
membrane without destroying the interactions of nuclear proteins.

We were already discussing the type and concentrations of detergents,
however we did not get final clue.

Can anyone help me out there with some suggestions or protocols or are
there good references (protocols) in the literature (for general
membrane dissolving and in particular nuclear membrane dissolving) ?

Thank you very much,

Werner



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  #2  
Old 02-16-2009, 06:23 AM
DK
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Default Detergent concentration for nuclear membrane dissolving

In article <[Only registered users see links. ].net> , "Werner Straube" <[Only registered users see links. ].de> wrote:

There isn't a detergent that does not interfere with all relevent
protein-protein interactions! So no matter what your friend will have
to play. That said, couple suggestions to start:

1. sodium cholate is a mild and dialysable detergent that at 1%
dissolves nuclear membranes quite well.
2. octyl glucoside is very mild and dissolves a lot of nuclear
proteins at 0.5-1% in thew presence of >200 mM NaCl.
3. heck, some interactions are not affected to a great extent by
0.1% SDS

DK
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  #3  
Old 02-16-2009, 02:18 PM
Dr Engelbert Buxbaum
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Default Detergent concentration for nuclear membrane dissolving

Am 15.02.2009, 16:29 Uhr, schrieb Werner Straube <[Only registered users see links. ].de>:


In my experience positively charged detergents like CTAB can dissolve
nuclei very well (better than SDS) while at the same time are generally
mild (milder than OG or DM) to proteins. Whether or not the interaction
that your friend is interested in is maintained under these conditions
he/she will have to find out.
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concentration , detergent , dissolving , membrane , nuclear


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