Re: BCL6

From:Kappeler Andreas

Hi Elaine
BCL6 is indeed a pain. We are using clone PG-B6p from Dako at a
concentration of 24 ug Ig/ml after HIER in a pressure cooker, 10 mM citrate
buffer, pH 6.0 (gave somewhat better results than an urea containing high pH
buffer). After looking at numerous follciular lymphomas and tonsils it is
our impression that good staining depends largely on quick fixation, as you
will regularly find stronger staining in the periphery (1-3 mm) of the
samples (where formaldehyde gets quickly) than in the centre (which may be
completely - and falsely - negative, as formaldehyde gets there only after
hours, if the sample is not sliced). So again a case to blame the whole
misery on preanalytic sins and errors ... However, even with such cases you
can get a wonderful, strong nuclear reaction if you are using biotinyl
tyramide amplification (Tyramide Signal Amplification TSA from NEN Life
Science products or Catalyzed Singal Amplification CSA from Dako (same stuff
as TSA; licensed from NEN)). Good luck and Happy Holidays!

Andi Kappeler
Institute of Pathology, University of Bern, Switzerland

-----Ursprüngliche Nachricht-----
Von: Elaine Dooley 
Gesendet: Donnerstag, 20. Dezember 2001 19:59
Betreff: BCL6

> Dear Histonetters,
> Has anyone had success staining formalin fixed paraffin embedded sections
for BCL6.  I have tried heat induced epitope retrieval with Trilogy (Cell
Marque)and high ph target retrieval  from DAKO on tonsil sections.  I have
tried both companies antibodies.  The best staining I had was with high ph
target retrieval and the antibody at 1:10.  But I need to get stronger
staining.  I have tried amplification on a Ventana immunostainer and also
tried staining by a manual method and it is just not dark enough.
> Would appreciate any suggestions
> Elaine Dooley HTL
> Gainesville Fl
> 352-265-0111 ext 4-4935
> Thanks in advance

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