I've done this before.  I just filled a prep kit with a the
biotin-conjugated secondary against goat (I used one from miltinyi and I
think the dilution I used was 1:200), then I photocopied the bar code
from Ventana's biotinylated secondary, put it on the prep kit, filled it
up, and replaced  Ventana's secondary.  I ran it by alone since that
secondary wouldn't pick up a signal from my normal mouse and rabbit
antibodies.  

 




Mark Adam Tarango HT(ASCP)

Histology/Immunohistochemistry Supervisor

Nevada Cancer Institute

One Breakthrough Way

Las Vegas, NV  89135

mtarango@nvcancer.org

Direct Line (702) 822-5112

Fax (702) 939-7663

  

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sebree
Linda A.
Sent: Wednesday, February 28, 2007 1:09 PM
To: Histonet
Subject: [Histonet] Secondary antibody for Ventana detection kits

Hello,

I have the need to optimize a goat anti-human antibody on the Ventana XT
immunostainer.  In order to incorporate an anti-goat secondary I am
using their "open secondary" protocol with an iView DAB detection kit.
I am wondering whose antibody people are using in these circumstances.
Ventana says that people have used Jackson ImmunoResearch's biotinylated
R x G F(ab')2 (cat. #: 305-066-003) for this purpose with success.  I
would like to actually talk to anyone who has done this as I have
concerns about their storage requirements and putting this antibody in a
dispenser.

Thank you in advance for your help,

Linda Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-3224
600 Highland Ave.
Madison, WI 53792
(608)265-6596
FAX: (608)262-7174

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