[Cytometry] Opinions about planned experiment

Vanda Juranic vanda_juranic at yahoo.com
Thu Feb 18 10:34:19 EST 2010


We have antibodies over several years old (some over 10!) and all are still working in the same concentration as when they were purchased with no observable loss of binding/intensity/whatever.

 Vanda Juranic Lisnic
Department of Histology and Embryology 
Faculty of Medicine, University of Rijeka 
B. Branchetta 20 
51000 Rijeka 
CROATIA 
Phone: +385 51 651 170 
Fax: +385 51 651 176




________________________________
From: Mario Roederer <roederer at drmr.com>
To: Florian Uhle <Florian.Uhle at chiru.med.uni-giessen.de>
Cc: cytometry at lists.purdue.edu
Sent: Thu, February 18, 2010 3:32:21 PM
Subject: Re: [Cytometry] Opinions about planned experiment

In general, it is better to rely on data than on opinion when designing experiments.  The data show that freezing is not good for phycobiliprotein conjugates of antibodies.  In addition, we (and many others) have stored antibodies at 4C for much longer than this without noticeable loss of intensity. 

As long as you are using saturating concentrations (titrate your antibody!), you should be fine.  Incidentally, your concern is the main reason that you should use an experimental staining control that you know is identical with each experiment.  It will help identify if your reagent  or procedures are variable.

mr

On Feb 18, 2010, at 9:07 AM, Florian Uhle wrote:

> Dear community,
> 
> thanks a lot for your great input within this short time. 
> 
> Some of you recommended not to freeze the conjugated antibody. My problem is, that it will take approximately 18 month to enclose the necessary number of patients. In my opinion, this is too long for 4 degree storage without loss of intensity. Any suggestions how to deal with this issue?
> 
> I also realized my lack of proper instrument QC and after browsing some manufacturer homepages, I believe that Spherotech's Calibration kit will be the solution to this. Does it make sense, to first perform automatic BD CaliBRITE /FACSComp compensation and afterwards the QC check using the Spherotech beads? 
> 
> All the best,
> 
> Florian
> 
> _____________________________________________
> 
> Florian Uhle
> Justus-Liebig-Universität Gießen
> Abteilung für Anästhesie, Intensivmedizin und Schmerztherapie
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> 
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