Cytokine measurement in transfected cell line

From: Jan Grawé (jan.grawe@rudbeck.uu.se)
Date: Fri Nov 22 2002 - 04:03:23 EST


Dear All,

I have a user who wishes to detect cytokine production in a
transfected cell line, see below . Use of at least one commercial
permeabilisation kit has failed.
Literature and email archive searches have yielded virtually no
information, so any insights are appreciated!

Jan Grawé

>Hi,
>
>I am trying to develop an intracellular staining protocol to detect
>intracellular protein in a tumor cell line
>by flow cytometry.  I have checked the literature and the technique
>seems to be used almost entirely for
>lymphoid cells.  Does this mean that other types of cells are
>difficult to permeabilize?
>
>Does anyone have any experience detecting intracellular protein
>within tumor cells using flow cytometry?
>
>I am transducing the cell line MB49 (murine bladder carcinoma) with
>recombinant adenovirus expressing IL-15,
>and I would like to detect IL-15 by intracellular staining.
>
>I would really appreciate any insight and/or comments.
>
>Thanks,
>Christina Ninalga


--
--------------------------------
Jan Grawé
Cellanalyslab
Rudbecklaboratoriet/C5
Uppsala Universitet
Dag Hammarskjölds väg 20
75185 Uppsala

tel: +(0)18-4714656
mobil:+(0)70-2577874
fax: +(0)18-552739
epost: jan.grawe@rudbeck.uu.se
http://www.rudbeck.uu.se/cellanalys



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