How stable biotin-avidin?

Paul N Hengen pnh at
Mon Mar 11 13:24:17 EST 1996

Zophonias O. Jonsson (zjons at wrote:

> I have noticed a number of non-isotope DNA/RNA hybridization techniques
> that use biotin-linked probes. How stable are the biotin and avidin
> proteins to high temperatures used in nucleic acid
> hybridization/dissociation, e.g. 65 degrees or higher. Secondly, how stable
> is the biotin - avidin interaction to higher temperatures? 

: The Biotin-avidin interaction is VERY strong.  (Unfortunately I don't
: have the binding constants at hand).  Biotin (vitamin H) is not a protein
: and is quite resistant to heat. Avidin is quite robust too.

The dissociation constant is on the order of 10^-12 so there is almost
no way to separate them without boiling in DMF. Hybridization or PCR
is commonly used with biotinylated DNA probes.

Paul N. Hengen, Ph.D. (pnh at
National Cancer Institute--FCRDC--Frederick, Maryland 21702-1201 USA
phone:(301) 846-5581 fax:(301) 846-5598
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