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SLTJP at CC.USU.EDU SLTJP at CC.USU.EDU
Tue Jan 24 23:03:36 EST 1995


>Greetings,
>
>        I have recently puified the bovine serum protein hemopexin.  I
>havan
:AP or streptavidin::HRP to
>bind the labelled protein.  The other possibility is that binding does
>occur, but the hemopexin::biotin/streptavidin::AP-HRP complex somehow
>impairs the function of the AP or HRP.  The streptavidin::AP-HRP is
>active when it binds transferrin::biotin.
>        Has anybody else had this problem?  Do you have an
>explanation/solution?
>        I am now trying to directly conjugate hemopexin to AP and HRP as
>an alternative method.  I would appreciate any insites you may have?
>
>P.S.  Has anybody used that radio-labelling kit from Amersham that uses
>[35-S] instead of [125-I]?  How does it work?
>
>Thanks, Reno
>
I have been doing some biotinylation of antibodies.  The biotinylation
has been working.  To check the available biotins, as opposed to the
total number of biotins bound Pearce has a relitivly cheap and very
easy HABA kit avalilable.  To modify the kit for avaliable biotins
do not degrade your protein of interest.  This way only the available
biotins react.  Hope this is of use.  Mick 

Mick Blake
SLTJP at cc.usu.edu
Utah State University
Dept Food Sciences
801 797 2123



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