intracellular interleukin, please help

From: andreas.simm@medizin.uni-halle.de
Date: Wed May 02 2001 - 07:53:31 EST


Hello again,
3 weeks ago, I tried to get some information about the analysis of
intracellular interleukins in monocytes. A lot of people asked me to
forward the answers to them, but until now I got only one answer
(thanks to Joost). Therefore I want to repeat my questions again
and forward my original mail.
We would be happy about every help.

Andreas

------- Forwarded message follows -------
From:			Self <andreas.simm@medizin.uni-halle.de>
To: cyto-inbox
Subject:		intracellular interleukins in monocytes? IL-18 antibody?
Date sent:		Thu, 12 Apr 2001 16:27:38 +0200

Hello everybody,
we want to analyse the expression of
interleukins in monocytes.

During the first analysis, a lot of
questions appeared. Therefore, I asked
some people and got no really clear answers.

Hopefully, somebody can help me.

1) We want to stimulate whole blood for up
to 24 hours and analyse the monocytes afterwards.
In which sort of tubes should we keep the blood
or is it possible to block the attachment of the
monocytes to the tube material?

2) If the monocytes are attached to the wall, is it
possible to include them in an analysis or are all
of them stimulated by the attachment per se?

3) We see that one have to predilute the blood in
RPMI or AIM-V medium before one stimulate the
cells. Which medium is the better one and why
do one have to dilute it?

4) Which heparin blood is the better one, Li-heparin or Na-heparin?
I see the question from yesterday and hope
to get the answer as well.

5)  Do anybody know a source of IL-18 antibodies
for intracelluilar staining?

Thanks for your help in advance

Andreas Simm

------- End of forwarded message -------


Andreas Simm
Klinik fuer Herz- und Thoraxchirurgie
Universitaet Halle
Ernst-Grube-Str. 40
06120 Halle

Tel.: +49 (0) 345 557 2647
FAX:  +49 (0) 345 557 2782



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