Phage display

Troels Wind wind at biobase.dk
Wed Jan 8 10:05:39 EST 1997


Mozart de Azevedo Marins (mdamarin at usp.br) wrote:
: Hi there,
:            We are interested in our lab. to ue the Phage display kit from 
: New England Biolabs in order to find a potential peptide ligands to a 
: rpotein we work with. I would like to know if any of you has experience 
: with their kit. And also I would like to know how long has to be the 
: target coated to the plate where the potential peptide ligands will be 
: exposed.

I havent used the NEB-system, but with phage displayed antibody libraries
the antigen (target) is usually coated over night at 4C.

To save yourself some problems I suggest that you optimise your coating
conditions. If you have an Ab against your target this can be done by
ELISA. Try varying the coating buffer (pH, ionic strength etc.) and the
plate used. We have used Maxisorb from Nunc (Denmark, NO CONNECTION) with
success.

In one instance we found that 10-20% glycerol in the coating buffer
was beneficial.

Hope this helps...

Troels Wind, M.Sc.
University of Aarhus
Institute of Molecular and Structural Biology
Denmark



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