In my experience with trying to do alk. phos. on ffpe decalcified bone for
osteoblasts the enzyme reactivity did not survive.  I had to do it on bone
embedded in GMA without decal.
Patsy

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of LORALEE
GEHAN
Sent: Tuesday, April 27, 2004 2:05 PM
To: mab70@medschl.cam.ac.uk; brucea@unimelb.edu.au;
histonet-bounces@lists.utsouthwestern.edu;
histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Re: HELP
PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee &Peggy/Tony, SOMEONE....)



   With  my  experience it won't survive the processing.  Try it plastic.
   The processing is a bit longer but the stain works nicely.

   Hope that helps.

   Loralee Gehan, HTL
   >From: Margaret Blount 
   >To: 'Bruce Abaloz' ,
   histonet-bounces@lists.utsouthwestern.edu,
   histonet@lists.utsouthwestern.edu
   >Subject: RE: [Histonet] Re: HELP
   PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee & Peggy/Tony, SOMEONE....)
   >Date: Tue, 27 Apr 2004 10:46:25 +0100
   >
   >Hi, Everyone,
   >
   >With  regard  to this problem, I was wondering if the alk phos enzyme
   would
   >survive  processing,  I  rather  think it would otherwise we wouldn't
   need to
   >add  levamisole  when immunostaining paraffin sections using alk phos
   as a
   >label. So why not process into wax then do the detection of alkaline
   >phosphatase and the other markers you wish to identify.
   >
   >What do other histonetters think? Is it worth a try?
   >
   >Margaret
   >
   >Margaret Blount
   >Chief Technician
   >Clinical Biochemistry
   >University of Cambridge
   >Addenbrooke's Hospital
   >Hills Road
   >Cambridge
   >CB2 2QR
   >
   >-----Original Message-----
   >From: histonet-bounces@lists.utsouthwestern.edu
   >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bruce
   >Abaloz
   >Sent: Tuesday, April 27, 2004 2:30 AM
   >To: histonet-bounces@lists.utsouthwestern.edu;
   >histonet@lists.utsouthwestern.edu
   >Subject: [Histonet] Re: HELP
   PLEASE(John/Gayle/Connie/Cynthia/Patsy/Lee
   >& Peggy/Tony, SOMEONE....)
   >
   >
   >Hi, my name is Danielle & I need some advice PLEASE -
   >I  have  been  using  alkaline  phosphatase  histochemistry to detect
   primordial
   >germ  cells  in  PFA-fixed  marsupial embryos. I use 1mg/ml Fast Blue
   salt and
   >1mg/ml Naphthol AS phosphate sodium salt in 0.2M Tris buffer, pH 9.4.
   The
   >staining  is  good  but  I  have  been told that it is not permanent.
   Ideally, I
   >would  like  to  dehydrate  these  stained  specimens,  embed them in
   paraffin and
   >then section and stain them immunohistochemically for other germ cell
   >markers  and  several  growth  factors.  Does  anyone have a protocol
   utilising
   >alkaline  phosphatase  histochemistry that would allow me to do this?
   Thanks
   >for any advice in advance,
   >
   >Dr Danielle Hickford
   >Research Fellow
   >Department of Zoology, University of Melbourne,
   >Australia 3010.>hickford@unimelb.edu.au
   >
   >--
   >BRUCE ABALOZ                           PH:61383446282
   >HISTOLOGIST                            FAX:61383447909
   >DEPT.of ZOOLOGY                        EMAIL: brucea@unimelb.edu.au
   >THE UNIVERSITY Of MELBOURNE.
   >VICTORIA.AUSTRALIA 3010
   > >
   >  >                Nobody  can  make  you  feel inferior without your
   permission.
   >-  Eleanor Roosevelt
   > >                         DANCE LIKE NO-ONE'S WATCHING
   >*********************************************************************
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   >--
   >BRUCE ABALOZ                           PH:61383446282
   >HISTOLOGIST                            FAX:61383447909
   >DEPT.of ZOOLOGY                        EMAIL: brucea@unimelb.edu.au
   >THE UNIVERSITY Of MELBOURNE.
   >VICTORIA.AUSTRALIA 3010
   >
   >                Nobody  can  make  you  feel  inferior  without  your
   permission.  -
   >Eleanor Roosevelt
   >                          DANCE LIKE NO-ONE'S WATCHING
   >*********************************************************************
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