Protein or peptide for Ab production?
ekuusist at JALUS.UKU.FI
Tue Nov 24 06:43:10 EST 1998
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.
University of Kuopio
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