IUBio

?Coating efficiency of an ELISA plate?

Pete bss3pk at surrey.ac.uk
Fri Nov 22 07:15:00 EST 1996


Zafer Altin <zafer at abconcepts.com> wrote:

>You may try coating test plates with biotinylated antigens followed by
>back-coating with some blocking buffer (e.g. 1% BSA plus Tween 20).
>Then add an optimized concentration of strepavidin-HRP to generate
>a readable signal(OD 1.0-2.0) when substrate added.  This will not give
>you an absolute amount of bound antigen but a fairly reasonable
>comparison among three antigens you anticipate to use. You should also
>determine biotin/collagen molar ratios to determine how similar
>the capacities of these three antigens to bind strepavidin-HRP are. 
>Good luck.

... ah, but can you be sure that all the collagens will biotinylate
with the same efficiency?

Pete

>Z. Altin
>Antibody Concepts
>zafer at abconcepts.com
>web site:http://www.abconcepts.com





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