[Histonet] re: Endogenous biotin blocking

[Tyrone Genade]

From: Sally Price <sprice2003 <@t> gmail.com>
Subject: [Histonet] Endogenous biotin blocking

> Its been my understanding that blocking is only necessary when one is certain
> that background staining is caused by endogenous biotin, but maybe I'm
> off-base here.  I look forward to eveyone's input.

I have been working on brain sections and without first blocking the
endogenous biotin I get very poor definition staining in sections and
no discernible staining in whole mounts. Any tissue that is
metabolically active will have large concentrations of biotin as it is
a co-enzyme in many of the enzymes involved in carboxyl-group transfer
(such as in glycolysis , the CAC and fatty acid metabolism).

The simplest way to see if you need to block is to do the experiment
with and without blocking. More than likely, you will need to block.
This is my opinion as a biochemist not a veteran histologist (which I
am not).

Hope this helps.
-- 
Tyrone Genade
http://tgenade.freeshell.org
email: tgenade <@t> gmail.com
tel: +27-84-632-1925 (c)
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