Why not use an avidin-biotin blocking kit (like those from Vector or Dako for example). They work well, are optiimized already and aren't that expensive.


-----Original Message-----
From: Tyrone Genade 

Date: Mon, 20 Oct 2008 16:07:07 
To: 
Subject: [Histonet] getting rid of biotin-avidin background


Hi all,

I have a problem and think I have figured out a solution to it but
need some expert advise.

I'm using biotinylated GS isolectin to stain frozen brain sections for
microglia. The GS isolectin is working very well but the
avadin-fluourophore is binding to the endogenous biotin in the tissue
causing a lot of back ground. I'm thinking of fist blocking the
sections by incubating it with unlabled avidin and then following it
with another blocking step with unlabled biotin so that all the
endogenous biotin is bound to avidin and the un-endogenous biotin
bound sites of the avidin are then bound to unlabled biotin. This
would then be followed by the GS Isolectin incubation etc...

Can anyone suggest a reasonable starting concentration for the first
biotin and avidin blocking steps? Do you think this will work at all?

Kind regards

-- 
Tyrone Genade
http://tgenade.freeshell.org
email: tgenade@freeshell.org
tel: +27-84-632-1925 (c)
********************************************************************************
"For there is one God, and there is one mediator between God and
men, the man Christ Jesus, who gave himself as a ransom for all."
        1 Timothy 2:5-6

_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet