[Histonet] Re: HELP PLEASE(John/Gayle/Connie/Cynthia/Patsy/Le e & Peggy/Tony, SOMEONE....)

[Margaret Blount]

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 <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Bruce
Abaloz
Sent: Tuesday, April 27, 2004 2:30 AM
To: histonet-bounces <@t> lists.utsouthwestern.edu;
histonet <@t> 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 <@t> unimelb.edu.au

-- 
BRUCE ABALOZ                           PH:61383446282
HISTOLOGIST                            FAX:61383447909
DEPT.of ZOOLOGY                        EMAIL: brucea <@t> unimelb.edu.au
THE UNIVERSITY Of MELBOURNE.
VICTORIA.AUSTRALIA 3010
>
>               Nobody can make you feel inferior without your permission.
-  Eleanor Roosevelt
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-- 
BRUCE ABALOZ                           PH:61383446282
HISTOLOGIST                            FAX:61383447909 
DEPT.of ZOOLOGY                        EMAIL: brucea <@t> 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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