[Histonet] DAKO ER and PR antibodies

[Dana Settembre]

Hello Theresa,
I use the same ER and PR antibodies that you use and I also use Dako's
Target Retrieval Solution
My method works well:
TRS for 40 minutes in a steamer
ER used at 1:20 and incubated for 30 minutes (more can't hurt, up to 60
minutes)
PR used at 1:30 and incubated for 20 minutes (more can' hurt either)
I use Dako's LSAB2 kit (which is less sensitive than your Envision +, I
might go that route and dilute my Ab's)

Good Luck
Dana Settembre
University Hospital-UMDNJ
Newark, NJ


>>> THERESA ROHR <trohratnyackhospital <@t> hotmail.com> 10/2/2003 8:00:08
AM >>>
Hi out there in Histonet land:

I am having a problem working up dilutions for DAKO's ER (M7047) and PR

(M3569) antibodies.
I am using the Envision+ Detection system

Suggestions from DAKO were:

ER  1:100 to 1:150 with Envision+ and a 20 minute target retrieval
PR  1:200 to 1:400 with Envision+ and a 20 minute target retrieval.

I've gone less and more and still cannot get any where near the
staining on 
these cases that went out to a IHC Laboratory than I am using as a 
comparison.

Any suggestions would certainly be appreciated.

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