[Histonet] RE: PCNA on cryosection

[C.M. van der Loos]

   Hi Renee,

   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

   Meibergdreef 9

   NL-1105 AZ Amsterdam, The Netherlands

   ----- Original Message -----
                                From 
   "Till, Renee" <TillRenee <@t> uams.edu>
                                Date 
   Wed, 23 Mar 2005 07:49:56 -0600
                                  To 
   histonet <@t> lists.utsouthwestern.edu
                             Subject 
   [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
   actually
   have  a specific stain in mind. One of my other Zymed kits says to fix
   in
   acetone,  so  I thought I might do that. Should you fix after cutting,
   or
   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
   1120 Marshall
   Little Rock, AR