>Hi
>
>I need to visualise CD31 in rat tissue. There appears to be only one
>monoclonal available from Research Diagnostics. It requires zinc fixation
>prior to paraffin embedding and  alk. phos detection.
>
>Does anyone have an opinion on
>1. Zn fixation
>2.what is the optimal chromogenic substrate for alk phos
>3. Dako mentions an endogenous alk phos inhibitor, levamisole which they
>put in some of their kits
>
>Thanks
>
>
>_____________________________________________________
>
>Nazlia Samodien
>
>Cardiovascular Research Unit
>Dept. of Cardiothoracic Surgery
>University of Cape Town Medical School
>Anzio Rd, Observatory, 7925, South Africa
>
>Tel. +27 21 406 6398 (office)
>        +27 21 406 6476 (Secretary)
>Fax + 27 21 448 5935
>
>
>Email: ctssamodien@samiot.uct.ac.za

***********

Dear Nazlia,

1. Recently there were some reports about zinc fixation on histonet!
2. One of the best AP chromogens is DAKO Fast Red. It provides you with a
very intense an crisp staining. A nuclear counterstain in blue with
haematoxilin gives a very nice contrast! Don't forget to mount up your Fast
Red stained sections aquously, as it will dissolve in alcohol and/or
xylene.
Alternatively use BCIP/NBT in dark blue/black. This chromogen provides more
sensitivity than Fast Red, although you probably don't need that for
detection of CD31.
3. As far as I know levamisole is not inside the kit. You have to put it in
yourself. The use of 3 mM levamisole is only useful for cryostat sections,
since endogenous alk. phosph. does not survive formalin-fixation and the
paraffin-embedding.

Have a happy staining day,
Chris