Re: [localization within lymphocytes]

From: astacchini (astacchini@molinette.piemonte.it)
Date: Wed Sep 25 2002 - 02:43:02 EST


We obtain good results for cytoplasmics and nuclear protein investigation in
human lymphocytes using the FIX and PERM kit of Caltag.
If you want to use a domestic assay you can fix the cells with  200 ul of 2%
paraformaldehyde for 10'in ice, wash with PBS , than permeabilize adding to
the pellet 50ul of 1% saponin in PBS for 5'in ice.
alessandra Stacchini

On Mon, 23 Sep 2002 19:55:59 +0200 (MEST), Lars Brichta wrote
> Dear all,
>
> is it possible to determine a protein in lymphocytes (isolated from whole
> blood) which is localized in the cytoplasmatic space and the nucleus
> as well? I think probably it would be necessary to fix the cells and
> to use a permeabilization agent which acts on both the cell membrane
> and the nuclear membrane as well?
>
> Does anybody know if there is a paper which gives any hints for such
> a method or the use of a kit?
>
> Thanks a lot!
>
> Lars Brichta
> Institute of Human Genetics
> University Clinics Bonn, Germany
>
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