[Histonet] RE: PCNA on cryosection
C.M. van der Loos
c.m.vanderloos <@t> amc.uva.nl
Wed Mar 23 12:42:12 CST 2005
PCNA isn't working very well on frozen tissue sections. At least not
in my hands some time ago. We use the monoclonal anti-ki67
(LabVision/Neomarkers) for human, mouse and rat instead.
If you wish to stain nuclear antigens in a cryostat tissue section you
better avoid acetone as a fixative as this will result into a quite
diffuse staining. My tip for a crisp nuclear staining on frozens is
just a 5 minutes fixation with NBF. No antigen retrieval is needed.
Hope this helps.
Chris van der Loos, PhD
Dept. of Pathology M2-230
Academical Medical Center
NL-1105 AZ Amsterdam, The Netherlands
----- Original Message -----
"Till, Renee" <TillRenee <@t> uams.edu>
Wed, 23 Mar 2005 07:49:56 -0600
histonet <@t> lists.utsouthwestern.edu
[Histonet] PCNA on cryosection
Can anyone give me a general protocol for what to do with cryosections
that will be used for the PCNA stain? I have the PCNA kit from Zymed.
They say you can use them on frozens, but that is about all the help
they give. I know I asked this question before, but this time I
have a specific stain in mind. One of my other Zymed kits says to fix
acetone, so I thought I might do that. Should you fix after cutting,
wait until you are going to use the slides? If you are going to wait I
would think the slides should be kept in -80? Any suggestions?
Renee' Till, HT
Arkansas Childrens Nutrition Center
Little Rock, AR
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