Help! Polyclonals to a polypeptide

Steve Dunn dunnsm at bbsrc.ac.uk
Mon Aug 7 06:05:55 EST 1995


How would I go about preparing a 70aa plant protein domain in order to
raise polyclonals? I have inherited a construct containing the domain
fused to an N-term. 6-His tag and the "smudges" observed on conventional
SDS-page gels are evidence of it's overexpression (yes!-I know I should
run Tricine gels for polypeptides of this size).  So, if I go with this
construct and QIA-Ni purify the fragment, what would I conjugate it to,
and how would I do this? Do I need to conjugate?  Alternatively are there
tried and tested smarter stragies I could employ (I'm quite happy to make
new constructs/fusions etc). Any advice would be much appreciated.

Steve 


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