[Histonet] re: problems with BD PECAM-1 / CD31

[Patsy Ruegg]

CD 31 in my hands requires enhanced detection systems as Jason uses with the
ABC elite I have also gotten good results with the DAKO CSA II Tyramide
detection system, not much luck with non-enhanced systems such as LSAB or
even Labelled Polymer.
Patsy 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of PALMER Jason
(SVHM)
Sent: Thursday, May 05, 2005 10:49 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] re: problems with BD PECAM-1 / CD31


Hi.

In response to Jackie O's and Melissa Gonzalez's comments on BD CD31 IHC
problems on FFPE tissues, I have been getting very nice staining recently
using a protocol developed with tips from Carrie Kyle Byrne, after i made a
similar query to histonet some time ago. Mostly use it on our Dako
autostainer, but works fine on the bench too.

Tissues fixed O/N in 4% PFA (Zn fixation not necessary) processed to
paraffin. H2O2 block 5 min, Dako proteinase k 8min RT, Dako protein block 30
min, primary 1:100 in Dako diluent 60 mins RT, secondary Dako rabbit anti
rat biotin 1:300 30 min RT, vector ABC elite 30 min RT then DAB 5 min. Gives
consistently good vascular staining for me, in connective and adipose rich
mouse tissues. I think the ABC elite is the key.

And i'm going to give the Santa Cruz PECAM Ab that has been talked about
recently a try too. Has been sitting in our fridge for a while as a second
option, and though don't really need it at the moment, thought i'd take it
out for a spin to see how it performs...

Hope this helps,

Jason

Jason Palmer
Bernard O'Brien Institute of Microsurgery
42 Fitzroy St, Fitzroy Victoria 3065
Australia
tel +61 3 9288 4018
email: palmerj <@t> unimelb.edu.au