Hi,
    It really sounds like she needs to dilute the antibody out further.
Usually even prediluted antibodies can / should be diluted. You could
probably cut down the incubation time too if you did the staining at room
temperature. We incubate for 30 minutes with good staining. Run a few titers
and I'm sure it will clean up.
Amos Brooks
----- Original Message -----
From: "Cindy Chard-Bergstrom (by way of Histonet)" 
To: 
Sent: Thursday, September 20, 2001 3:56 PM
Subject: Re: ER staining


> Good afternoon to all,
> I am inquiring on behalf of one of our graduate students. She is trying to
> do ER staining on canine tissue. To date she has tried staining uterus and
> has used human uterus as a positive control. The problem she is having is
> with background staining and excessive staining of cells in the stroma and
> myometrium which is as strong or stronger than the epithelial area that
she
> is trying to stain. This is not an antibody that I have ever used. Any one
> out there know any tricks for this antibody? Current protocol includes a
> prediluted antibody, heat retrieval using a steamer, overnight incubation
> at 4 degrees C and room temperature. Detection is by ABC, chromogen is
DAB.
> All suggestions are appreciated. Thanks to all.
>
>
>
> Cindy Chard-Bergstrom B.S.,HT(ASCP),QIHC
> Microbiologist III
> Kansas State University
> chardb@vet.ksu.edu
>
>
>