[Histonet] (no subject)

[Rene J Buesa]

Souad:
  I used CD31 (placenta as + control) from Dako (mouse monoclonal) at 1:20 on formalin fixed paraffin processed tissue and HIER citrate at pH6; primary during 30 minutes. Always with good results.
   The difference with your protocol is that you dilute more but at the same time you incubate overnight and this should compensate for your 10 times more dilution rate.
  I don't really see anything wrong with your protocol unless something happens during the overnight incubation; too low temperature perhaps?
  I would try first a shorter incubation and less dilution.
  Hope this will help you.
  René J.

"Belmadani, Souad" <sbelma <@t> lsuhsc.edu> wrote:
  
Hi,
im trying to stain endothelium in mice hearts, i used the anti CD31 from abcm and it did not work , now im using the FVW from DAKo again its not working. im using the antibody at 1;200 dilution overnight and no staining at all 
its formalin paraffine section and im using the citrate buffer ph 6 as antigen retrieval ....
any ideas why its not working 
i apprecite your help
thanks 
souad
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