Protein or peptide for Ab production?

Erkki Kuusisto ekuusist at JALUS.UKU.FI
Tue Nov 24 06:43:10 EST 1998


Hi netters,

What is the method of choice to prepare an antigen for antibody 
production, starting from cDNA sequence? I have the cDNA of a protein 
for which I would like to make Westerns and, possibly, 
immunohistochemistry. A commercial Ab is not yet available. The 
protein is from rat, 55 kD, likely soluble and cytoplasmic, and 
contains several putative phosphorylation sites.

Is it best to express in bacteria the protein with a peptide tag 
sequence, and affinity purify the tagged protein? If so, which 
purification tag would be most practical?

Or, for best results, am I better off using a synthetic peptide as 
the antigen instead of the complete protein?

Any suggestions would be helpful. Thank you in advance.

Erkki Kuusisto
University of Kuopio



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