Anomaly in biotin tagged peptide. What could be the reason?

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Adam B Shapiro

The streptavidin monomer is about 10 kDa, so I would guess that you are seeing the undissociated biotin-streptavidin monomer complex.

Prajakta Tathe

Thanks Adam. I want to do in vitro ubiquitination assay using this peptide as a substrate. I observed that the full-length protein is getting ubiquitinated but not this peptide. Does biotin tag is masking the peptide?

It may also be that the peptide is not large enough, or lacks sufficient determinants, to be recognized by the ubiquitin ligase.