[Histonet] Re:Endogenous biotin blocking

Tyrone Genade tgenade <@t> gmail.com
Thu Sep 3 05:01:41 CDT 2009


> From: "Hobbs, Carl" <carl.hobbs <@t> kcl.ac.uk>
> Subject: [Histonet] Re:Endogenous biotin blocking

> If you are in  a research lab and find that you will need to apply biotin
> blocking regularly, you can make up your own avidin/biotin solutions  if
> costs are a major factor.

To overcome the problem of endogenous biotin I incubated the sections
in an avidin solution. The avidin was the avidin-DH form that comes
with the Vector Stain ABC kits. I added it to my blocking solution as
per the manufactures instructions and incubated it on the sections for
1 hr at room temperature. There after the sections were washed 3x in
buffer and then I added biotin to a final concentration of 1 mg/mL to
the blocking solution and incubated for 15 minutes at room
temperature. After the biotin step the biotinylated probe in blocking
solution was added and incubated over night at 4 deg C.

This worked very well! Almost all back ground due to the endogenous
biotin was quenched.

Tyrone Genade
email: tgenade <@t> gmail.com
tel: +27-84-632-1925 (c)
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