AW: [Histonet] double staining on the ventana XT

[Gudrun Lang]

Kim,
We also use the Basalcellcocktail (with p63).
The protocol is put together of the two single protocols for the single
markers with the usual incubation times. The first HIER step is naturally
the retrieval for both antigens. The first part is the dab-kit. After the
colour-development a second HIER step takes place. You can choose time and
temperature (eg. 60 degrees, 15 min; or 90 degrees 8 min). That depends on
how easily your first primary and secondary antibodies can be denatured.
Than the Red-kit follows. At the end is counterstain and blueing.
If the first primary and sec. ab are'nt denatured you get an overall red
result. If the second HIER step is too harsh, you see it in the morphology
or high background.
Gudrun


-----Ursprüngliche Nachricht-----
Von: Artim, Kimberly [mailto:ArtimK <@t> slhn.org] 
Gesendet: Donnerstag, 22. Jänner 2009 18:02
An: gu.lang <@t> gmx.at
Betreff: RE: [Histonet] double staining on the ventana XT

Guadrun,
 Would you be able to provide me more detailed protocol information for the
prostate biopsy staining.  Are you using P63 as well?  Any information you
provide would be greatly appreciated.
 Thank you, 
Kim

Kimberly Artim, HT (ASCP)
Technical Coordinator, Anatomic Pathology
St Lukes Hospital & Health Network
801 Ostrum Street
Bethlehem, PA 18015
610-954-4832


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gudrun Lang
Sent: Thursday, January 22, 2009 11:57 AM
To: histoinfo <@t> comcast.net
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: AW: [Histonet] double staining on the ventana XT

Hi Jennifer,
we perform the doublestain routinly. The preferred protocol is CK HMW/p504s
on prostata biopsies. 
The protocol is easy to establish and the staining results are very good.
The run lasts about 4 hours and we do it overnight. You need the Red and the
DAB ultraview kit. No extra reagens. 
Gudrun

-----Ursprüngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von
histoinfo <@t> comcast.net
Gesendet: Donnerstag, 22. Jänner 2009 01:41
An: Histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] double staining on the ventana XT


Greetings,
We are looking into doing some immuno double staining. I searched the
archives and found a few similar inquiries but no responses. Anyone with
experiences to share I would love to hear what you have to say. Everything
from what not to do to what works well for you.
Many Thanks,
Jennifer



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