I am fusing small peptides around 10 to 30 a.a into the C-terminus of a larger protein. The question is if I need to design specific antibodies against those small epitopes?
I actually have two purification steps, One for the larger protein for which I Use the specific antibody. However, for the second purification that includes my fusion proteins I need another antibody. As the small portion of the protein (as the antigenic epitopes) are fused, I was wondering how may I have an appropriate antibody which correspond to the epitopes I am using?
Do you recommend any special method or company for that? Or any idea of any alternative technique ?
Many thanks,
mah
Thanks Goran for the answer.
I actually fuse a small peptide from a human pathogenic virus into another viral capsid protein. The viral capsid protein includes his-tag and of course those small peptides will be tagged too. So first I did one transformation for the capsid protein and confirmed it with western blot. The next step is to fuse the small peptide of the human virus into my capsid protein. The question is how can I recognise the proteins which have the fusion protein from the one that are empty? Both have his tag. How can I confirm it using western or further down for purification? So, I thought that I probably need to design antibody for that specific peptide because I am not sure if the commercial available Ab against the virus of interest will recognise the peptide.
Thanks
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