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 <[EMAIL PROTECTED]> Date: Mon, 20 Oct 2008 16:07:07 To: <histonet@lists.utsouthwestern.edu> 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: [EMAIL PROTECTED] 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
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