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

[PALMER Jason (SVHM)]

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

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