[Histonet] double immuno staining
vandries <@t> vub.ac.be
Wed May 5 09:19:08 CDT 2004
I need some help.
I am struggling with a double immunostaining for somatostatin receptor and
All my previous double stainings worked beautifully, but they were performed
on fresh frozen acetone fixed pancreatic tissue.
For staining of somatostatin receptor I have to fix with formaldehyde. I use
20 minutes 0.75% PFA perfused tissue which is snap frozen embedded in
tissue freezing medium and cut on a cryostat. I was advised to do it this
way to prevent loss of antigenicity.
Morphology is quite good.
This fixation works for the somatostatin receptor staining, but it kills the
I was able to retrieve most of the desmin signal by EIER for 10 minutes with
trypsin-EDTA, but now the morphology is awful.
Can someone give me some advice?
Veronique Andriessen BAS
Lab. Molecular Liver Cell Biology,
Free University Brussels (VUB)Belgium
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