[Cytometry] Conjugation of antibodies with tandem dyes

McClellan, Steve smcclell at UFL.EDU
Tue Feb 10 12:01:28 EST 2009


Hi Julie,
I have been using the Lighting Link kits for a while now.  They really ARE as good as they claim.  The lack of need to purify after conjugation means no loss of antibody.  They are stable now in my hands up to six months.  The only thing I will say is that these are to be used for flow, where the excess dye is easily washed away.  I tried one on a tissue for confocal and the background was too high, so if you are labeling something for immunocytochemical staining, you will need to do a post labeling column purification.  I've done MAbs and other proteins with great success. Feel free to contact me if you have any specific questions. 
Kind regards,
Steve

Steve McClellan
Sr. Biological Scientist
Interdiciplinary Center for Biotechnology Research(ICBR)-
Flow Cytometry Core Laboratory
University of Florida
352-273-8185 (office)
352-273-8070 (fax)
smcclell at biotech.ufl.edu
Cancer/Genetics Research Complex-Room 292A


-----Original Message-----
From: cytometry-bounces at lists.purdue.edu [mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Parmentier, Julie
Sent: Monday, February 09, 2009 6:34 PM
To: cytometry at lists.purdue.edu
Subject: [Cytometry] Conjugation of antibodies with tandem dyes

Hello everyone!

I'm looking to conjugate some antibodies to tandems (Cy5-PE, Cy7-PE, and Cy7-APC), and was wondering if anyone has any experience with Innova Biosciences "Lightning-Link" conjugation kits.  I've visited Dr. Mario's site for his advice, and found a great protocol that would absolutely be my first choice, except who can resist "The World's Easiest Conjugation Kits".  I do love easy, and more than that, I love a kit.  But I am worried that "easiest" will not translate into "best", or even "good-enough".  So before I pin my hopes on this, has anyone tried it out, and if you have, how did it work?? Any tips/advice would be greatly appreciated..If no one has tried it out I'd be willing to take a shot at it, see how it goes,and let you know how it turns out.....Thanks everyone!


Julie Parmentier
Scientist
Cancer Biologics
AstraZeneca
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