[Histonet] RE: PAX-8 background staining Ultra platform

[Sebree Linda A]

We also use Ventana's PAX-8, manufactured for VMS by Cell Marque.  Our protocol on our Ultras is 36" CC1, 16" incubation @ 36 degrees C.

Linda A. Sebree

University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
600 Highland Ave.
Madison, WI 53792

(608)265-6596
FAX: (608)262-7174

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Vanessa Perez
Sent: Wednesday, June 05, 2013 8:47 AM
To: Obregon, Cecilia; histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: PAX-8 background staining Ultra platform

We use std cc1  
Antibody incubation 36 min @ 36degrees

WE use the PAX-8 that comes in the ventana dispenser clone MRQ-50...i think its actually a cell marquee one tho

Vanessa 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Obregon, Cecilia
Sent: Tuesday, June 04, 2013 4:34 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] PAX-8 background staining Ultra platform

Anyone running PAX-8 on the Benchmark Ultra succesfully?

I have tried numerous protocols and antibodies, and still not able to obtain good results. I'm getting some background/cytoplasmic staining especially in normal kidney along with nuclear staining in the distal tubules and collecting ducts. I can't seem to completely be able to get it of it without compromising the signal. I have tried the Cell Marque MRQ-50 clone, the Biocare BC12, and now trying the Polyclonal Proteintech ...which looks promising.

It's hard to duplicate the protocols that other institutions have in place mostly because the platforms are different. Our institution only has Ventana platform, and they're suggesting we try their new detection kit for this marker: Optiview. I can't honestly imagine buying a detection for just one antibody...

We currently run UltraView Detection.

Any inputs or suggestions would be greatly appreciated.

Cecilia M. Obregon
Memorial Regional Hospital
3501 Johnson Street
Hollywood, FL 33021
Ph: 954-265-5317
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