MAP peptides for Abs

Tom Doak tom_doak at hlthsci.med.utah.edu
Thu Apr 28 14:14:28 EST 1994


We hope to make antisera to synthetic peptides representing our favorite
ORF.  I had thought to do this by a fairly conventional route: have the
pep synthesised, conj to KLH with a Pierce kit, have it injected into
rabbits, and hope.  However the AppliedBiosystems MAP substrate for
peptide synthesis has been suggested, but not by anyone with much
experience using it.  So MAPs are Multiply Attacted Peptides, or some
such, giving a small tree of peptides, each branch is another of the
synthesied peptides.  The substrate comes with either 4 or 12 branch
sites.  Supposedly this is large enough to be used directly as an
immunogen,  without coupling it to a carrier.  Also you no longer have to
worry about the immune responce to the carrier.  So questions:
-  Do these actually make good immunogens? KLH is much bigger, and is
supposed to stimulate the immune system in addition.  
-  How much do I need to use per bunny per injection?
-  Can they be easilly coupled to Ab affinity columns, ELISA plates, or
nitrocellulose?
-  Would anyone tell me to rush right out a do it.
-  What am I overlooking entierly?

Thanks so very much,

Tom Doak
tom_doak at hlthsci.med.utah.edu
Cellular Viral and Molecular Bio. (CVMB)
Med Center, UofUtah
SaltLakeCity, Utah



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