I agree with Robin, it is true for simple, direct detection systems. But for indirect systems where you would be detecting using secondary antibodies or hapten conjugated fluors it may be more important to use isotype controls (compared to 'no antibody' contols)to rule out nonspecific binding of IgG or haptens (eg. avidin class, biotin etc.,) ragoo *********************************************************** Arumugham Raghunathan, Ph.D Group Leader, Flow Cytometry Group Cellular Analysis Section Molecular Staging Inc., 300 George Street, Suite 701 New Haven, CT 06510 Tel# 203-772-5074 Fax# 203-776-5278 http://www.molecularstaging.com ************************************************************* -----Original Message----- From: Robin Barclay [mailto:Robin.Barclay@ed.ac.uk] Sent: Thursday, May 23, 2002 3:25 AM To: cyto-inbox Subject: Re: controls I stopped running isotype controls in any established situations where it could be shown that they were no different from the no-antibody control (do you also run these?) - and so seemed like a waste of money, sample and time - hardly no-brain reasons. Robin Barclay PhD Lead Scientist / Honorary Senior Lecturer SNBTS Academic Transfusion Medicine Group John Hughes Bennett Laboratory University of Edinburgh Dept of Oncology Western General Hospital Edinburgh ----- Original Message ----- From: "Julie L Wieseler Frank" <frankjw@psych.colorado.edu> To: cyto-inbox Sent: Tuesday, May 21, 2002 4:35 PM Subject: controls > > hello all! > > okay, so when i was trained to run flow experiments, i was STRONGLY > encouraged to use isotype controls for each label i used to evaluate non- > specific binding. is this considered a highly conservative approach? i am > working with somebody who shrugs off my need/want to run isotype controls, > and i feel baffled. > > i appreciate feedback -- this seems like a no-brainer, but it would seem i > am missing something! > > -- > Julie L. Wieseler-Frank, PhD > Research Associate > Department of Psychology > Campus Box 345 > University of Colorado at Boulder > Boulder CO 80309-0345 > tel: 303 735 2295 > fax: 303 492 2967 > frankjw@psych.colorado.edu > >
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