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I would try Wheat germ agglutinin (WGA). It binds to glycosylated proteins found in the cell membrane and extra cellular matrix. It is commonly used for as cell surface marker. Its advantage is that tends not to be sensitive to fixation as antibody targets can be. You can purchase it conjugated with a range of fluorophores (eg 488, 568, 647 etc). Simply dilute the stock in PBS and incubate for couple of hours (try 1 in 50 dilution as a starting point). I typically include it with the secondary antibodies in my labelling runs.
Cheers,
David
David Crossman PhD
Biophysics & Biophotonics Research Group
Department of Physiology
Faculty of Medical and Health Sciences
The University of Auckland
Private Bag 92019
Auckland 1142
New Zealand
Phone (64)-9-3737599 ext 89964
Fax: (64)-9-373 7499 attn: D Crossman
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From: Confocal Microscopy List [[log in to unmask]] on behalf of Cromey, Douglas W - (dcromey) [[log in to unmask]]
Sent: Thursday, August 15, 2013 8:58 AM
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Subject: generic membrane marker for FFPE tissue?
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I am working with a lab that has been trying to use Cadhedrin as a generic membrane marker for formalin-fixed, paraffin sectioned rat liver. They are not having much luck. Is there a good fixed tissue membrane marker that works reliably on paraffin sectioned tissue that you can recommend?
Because of ongoing problems with autofluorescence, it would be highly preferable if the membrane fluorescence was not in the typical FITC/GFP emission range, we would highly prefer something more red-shifted.
Doug
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Douglas W. Cromey, M.S. - Associate Scientific Investigator
Dept. of Cellular & Molecular Medicine, University of Arizona
1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA
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