Maria,
 you need to block the 1st primary antibody with IgG, and then use
appropriate serum, before using another primary from the same species.
I would use a different primary AB from a different species, i.e. if u used
goat anti.. The 1st time (when mistake was made) now use rabbit anti..

If you have to use another primary from the same species - I suggest review
excellent protocols on jacksonimmuno.com (labeling with 2 primaries from the
same species). This works, although may give you some background if blocking
is not done properly.

Igor

Dr. Igor O. Nasonkin
Research Fellow
National Institutes of Health/NEI
9000 Rockville Pike, MSC 1864
Bldg 10, Room 10B11
Bethesda, MD 20892
Tel: 301-443-7398 ­work
      617-388-4104 ­cell
Fax 301-480-1769
email: nasonkini@mail.nih.gov
http://www.nei.nih.gov/intramural/nnrl.asp


------ Forwarded Message
> From: 
> Reply-To: 
> Date: Tue, 20 May 2008 03:16:16 +0000
> To: Linke_Noelle ,
> , "Mejia, Maria"
> 
> Cc: 
> Subject: Re: [Histonet] one heck of a IHC question regarding
> fixedfree-floating brain sections
> 
> histonet@lists.utsouthwestern.edu

------ End of Forwarded Message


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