From owner-cytonet@hgmp.mrc.ac.uk Thu Oct 2 13:29:45 2003 Return-Path: X-Original-To: cytonet-outgoing Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with SMTP id D247A7D1A6 for ; Thu, 2 Oct 2003 13:29:45 +0100 (BST) X-Original-To: cytonet-list@hgmp.mrc.ac.uk Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 58CDF7D211 for ; Thu, 2 Oct 2003 13:29:45 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 7FD747D1A6 for ; Thu, 2 Oct 2003 13:29:44 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.11.6+Sun/8.8.8) id h92CThF15362 for cytonet-list@hgmp.mrc.ac.uk; Thu, 2 Oct 2003 13:29:43 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: klump.hannes@mh-hannover.de ("Hannes Klump") X-Newsgroups: bionet.cellbiol.cytonet Subject: dsRed2 / GFP in primary cells Date: 2 Oct 2003 13:29:42 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 30 Message-ID: Mime-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: 7bit X-Complaints-To: news@net.bio.net X-Original-To: cytonet@net.bio.net X-Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id D3E567D211 for ; Thu, 2 Oct 2003 13:29:41 +0100 (BST) X-Received: from MailExt1.MH-Hannover.DE (thor.mhrz.mh-hannover.de [193.174.104.33]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 139FF7D1A6 for ; Thu, 2 Oct 2003 13:29:35 +0100 (BST) X-Received: from nanna.mhrz.mh-hannover.de ([172.20.1.99] helo=Mail.MH-Hannover.DE) by MailExt1.MH-Hannover.DE with esmtp (Exim 3.02 #1) id 1A52aM-0005Sg-00 for cytonet@net.bio.net; Thu, 02 Oct 2003 14:29:34 +0200 X-Received: from [172.20.117.118] (helo=PC3165922) by Mail.MH-Hannover.DE with smtp (Exim 3.02 #1) id 1A52aM-0007mL-00 for cytonet@net.bio.net; Thu, 02 Oct 2003 14:29:34 +0200 X-To: X-Priority: 3 (Normal) X-MSMail-Priority: Normal X-Mailer: Microsoft Outlook IMO, Build 9.0.2416 (9.0.2911.0) X-Importance: Normal X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2600.0000 To: cytonet@hgmp.mrc.ac.uk Sender: owner-cytonet@hgmp.mrc.ac.uk Precedence: bulk Hi out there! I have two questions for you regarding the use of dsRed2 (and GFP) in primary cells: first: did any of you experience any toxicity or strange effects of dsRED2 on such cells yet? I have the feeling that this protein does not behave "neutral" in hematopoietic cells, but leads to strange selective effects. 2.) when coexpressing GFP and dsRed2 - do you have any problems with compensation during FACS analysis? We are using a FACScalbur from BD and at least in my hands, dsRED2 (measured in FL2) heavily emits into the FITC channel (FL1). Any ideas? Cheers, Hannes -- Hannes Klump, Ph.D. Hannover Medical School Center of Internal Medicine Dept. of Hematology and Oncology Experimental Cell Therapy, OE5120 Carl-Neuberg-Strasse 1 30625 Hannover Germany Tel. (++49)511 532 9473 Fax.: (++49)511 532 4521 e-mail: klump.hannes@mh-hannover.de --- .