| | |||||||
| Register | Search | Today's Posts | Mark Forums Read |
| Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique. |
| | LinkBack | Thread Tools | Display Modes |
|
#1
| |||
| |||
| 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 |
|
#2
| |||
| |||
| 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 |
|
#3
| |||
| |||
| 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. |
| Tags |
| concentration , detergent , dissolving , membrane , nuclear |
| Thread Tools | |
| Display Modes | |
|
|
| | ||||
| Thread | Thread Starter | Forum | Replies | Last Post |
| composition of water and detergent | jsrsol | Protocols and Methods Forum | 0 | 06-23-2009 09:34 AM |
| Western Blot Mystery - Developed bands don't adhere to membrane | cyrus | Western Blot Forum | 4 | 01-20-2009 08:08 PM |
| Low concentration miniprep | Utsugi, Heidi K | Protocols and Methods Forum | 0 | 01-15-2009 05:16 PM |
| New Saccharomyces Sequences 07/30/03 | SGD Sequences | Yeast Forum | 0 | 07-30-2003 06:19 PM |