Garry,

I've got an idea. You are having a nuclear counterstain problem with
immunos.
Try to find out if the antibody your using is available as a DNA (or RNA)
probe.
If so, perform the test as a in-situ hybridization procedure.  That should 
help in deciding if the marker is cytoplasmic or nuclear.

Donald Awbrey, HT(ASCP), QIHC
Electron Microscopy / Image Analysis
Harris Methodist Hospital
Fort Worth, Texas


-----Original Message-----
From: Garry Ashton [mailto:GAshton@picr.man.ac.uk]
Sent: Tuesday, August 15, 2000 9:05 AM
To: 'Histonet@pathology.swmed.edu'
Subject: immunostainig


Dear all,
Has anybody out there had any experience of a haematoxylin counterstain
being prevented from working, after immunostaining with an antibody. The
researcher presenting the problem doesn't know whether the Ab is nuclear or
cytoplasmic marker.
It cannot be fixation or necrotic tissue causing the problem as the negative
controls apparently stain fine as does her staining with other antibodies.
Any suggestions are welcome.
Garry


Paterson Institute
Wilmslow Road
Manchester
M20 9BX
UK