Yes sorry my message was assuming your primaries were from different 
species, otherwise indeed you need to use specialized blocking 
methods ... I think I am too tired to be emailing clearly!!!



>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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