[Histonet] Help!!!

[Katri Tuomala]

Hi Valerie,
I would recommend at least trying the biotin block. Certain tissues (liver, 
kidney) are rich in endogenous biotin and although formalin fixation often 
masks it, the HIER will bring it back.
Katri

----- Original Message ----- 
From: <vsailes <@t> nd.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Wednesday, May 11, 2005 10:44 AM
Subject: [Histonet] Help!!!


> Hello Histonetters,
> I'm working on an IHC protocol for Albumin -- staining FFPE mouse tissues. 
> I'm
> new to working out conditons for antibody protocols and could use some 
> help.I'm
> testing the antibody at different dilutions and I appear to have positive
> staining but I'm also seeing small areas in the negative that shows 
> positive
> staining. Liver is my control and I use steamer (HIER) for 30 minutes, my 
> steps
> include a 30 min quench in 1.6% hydrogen peroxide, NS block, Antibody, 
> Secondary
> antibody(Biotin), Streptavidin, Chromogen, Counterstain
> My concern is that I'm using buffers that contain BSA and I'm wondering if 
> that
> is causing staining to appear or should I be using avidin/biotin block 
> along
> with a peroxide block. Any suggestion would be helpful. Thank you for your 
> help
> in advance.
> Valerie
>
>
>
>
>
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