[Histonet] 34BE12 on prostate[Scanned]

[Katri Tuomala]

Hi Jean,
Yes,  clone 34 BE12 can be a problematic one. Over the years we have 
struggled same as you, but seem to have pretty good handle on it now. Our 
antibody is same as yours and we find, that more intense HIER is required. 
We are using citrate buffer (EDTA or others may be better, you'll have to 
try) in Decloaker. Our citrate buffer is home made and we add Tween 20 into 
it. Our dilution of the primary antibody now is 1:30.
I hope this helps...
Katri

Katri Tuomala
Hamilton,Ontario, Canada

----- Original Message ----- 
From: "Jean Gillson" <jean.gillson <@t> elht.nhs.uk>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Friday, February 25, 2005 6:02 AM
Subject: [Histonet] 34BE12 on prostate[Scanned]


> Dear all fellow histologist,
>
> We seem to have a recurrent problem with 34BE12 IHC staining on both
> prostate chippings and prostatectomy specimens.  Some basal cells stain
> whilst others that should stain are negative.  Our procedure is Trypsin 
> for
> 12 minute at pH 7.8 and 37 degrees C.  We use a Dako autostainer machine 
> and
> our antibody is from Dako.  Previous spec sheets state Trypsinisation but
> our most recent one states heat retrieval.  So as a test we did both
> retrieval methods.  Oddly enough, microwaved in Dako retrieval solution 
> was
> slightly better in prostatectomy specimens than trypsin (still uneven
> staining) and worst in chipping specimens.  Can anyone offer any advice?
> What antibody panels are people doing for prostate cases?  Help 
> desperately
> needed!!
>
> Jean Gillson BMS2
>
> Histology Department
>
> Blackburn Royal Infirmary
>
> Blackburn
>
> UK
>
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