Hi
I want to test the binding of one peptide to another, but there aren't antibodies to either of these peptides I a m interested in. Is there a way to add a tag to my peptides within my lab without professional help? Are there any biotins available that will only bind to amine terminus on the peptide and not all amine groups so I can perform an ELISA.
I am not interested in GST or FLAG tags as I would need the peptides specially synthesized.
Thank you for your suggestions.
Your best bet is to have the shorter of the 2 peptides synthesized with the N-terminal biotin.
I don't know of a method for labeling the alpha-amino group to the exclusion of lysine residues. However, if there are no lysine residues in your peptide, then you can use a primary amine-reactive biotin analog (such as biotin isothiocyanate or biotin succinimidyl ester) to label the alpha-amino group selectively.
Can you suggest a positive control for the peptide-peptide interaction? All I find on the internet is peptide-protein interaction
It depends how you are going to do the experiment. One way would be to bind the unbiotinylated peptide to the plate, then probe it with the biotinylated peptide, followed by a streptavidin-HRP conjugate. The positive control could then be to bind the biotinylated peptide to the plate instead of the unbiotinylated one.
If your plan is to bind the biotinylated peptide to a streptavidin-coated plate and probe it with the second peptide, then the positive control would be to make the second peptide with a biotin tag and bind that to the streptavidin plate.
Of course, it didn't occur to me to do that! This would definitely work in our scenario. I think we will go ahead with biotinylated peptides since it would be the most straightforward option.
Sometimes you're looking for more complicated answers and forget to think of the simple solutions.
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