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A Monoclonal Antibody That Discriminates between SNAP-Tagged and CLIP-Tagged Proteins

: Bialon, M.; Grezella, C.; Friesen, L.; Sieben, T.; Pham, A.-T.; Fischer, R.; Barth, S.; Püttmann, C.; Stein, C.


Monoclonal Antibodies in Immunodiagnosis and Immunotherapy 35 (2016), Nr.3, S.141-147
ISSN: 2167-9436 (online)
Fraunhofer IME ()

SNAP-tag technology allows recombinant proteins to be covalently labeled to O6-benzylguanine (BG)-modified substrates with 1:1 stoichiometry. By attaching according fluorophores, this method is ideally suited for in vitro and in vivo imaging, as well as protein interaction analyses. Fluorophores modified with BG react with the SNAP-tag, whereas those modified with O2-benzylcytosine (BC) conjugate to a more recent derivative known as the CLIP-tag. The orthogonal substrate specificity of the SNAP- and CLIP-tags extends the range of applications by allowing double labeling. We previously developed a monoclonal antibody (mAb) that recognizes both tags. In this study, we describe a new mAb, which is specific for the SNAP-tag alone. Therefore, this mAb allows discrimination between SNAP- and CLIP-tags within a broad range of immunological methods, including enzyme-linked immunosorbent assays, western blotting, flow cytometry, and immunohistochemistry.