[Histonet] IHC for estrogen receptor beta

Hermina Borgerink hborgeri <@t> wfubmc.edu
Mon Jun 9 12:04:02 CDT 2008

Andi, I have succesfully used Novocastra's mouse monoclonal ER beta
(Clone EMR02) on both human and non-human primates. Pre-treatment with
Citrate buffer at pH 6.0 for 30 minutes using a Black and Decker rice
steamer, with a 20-minute cooling down period.  I dilute my antibody at
1:50 in Tris buffer containing 0.5% casein and Tween-20, and incubate
overnight at 4 degrees C.  I use an anti-mouse biotinylated secondary
(1:20) and a streptavidin-alkalinephosphatase (1:20) from BioGenex,both
for 20 minutes at 37 degrees C.  I use Vector Red substrate for
visualization, and counterstain with Mayer's hematoxylin. 


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Andi
Sent: Monday, June 09, 2008 12:41 PM
To: Histonet
Subject: [Histonet] IHC for estrogen receptor beta

Hi all
has anybody been successfully using mouse-anti hu estrogen receptor
beta, clone 14C8 (GeneTex) on FFPE tissue? We have tried 8 different
pretreatment variants on 6 different samples of breast carcinoma
(including normal mammary glands) at Ig conc of 4 and 8 mg/l.
Visualization with polymer/HRP and DAB. If you were successful, would
you mind sharing your protocol? If you were successful with a different
anti-estrogen receptor beta antibody, I would be interested, too (clone
designation, company)! Many thanks!
Andi Kappeler
Institute of Pathology, University of Bern, Switzerland
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