Targeted and Reversible Biotinylation with Biotin-SS-Tyramide

Published on 28/01/2016

Our new Biotin-SS-Tyramide linker is a valuable tool for peroxidase-promoted targeted protein biotinylation. By using peroxidase-tagged antibodies, proteins and protein clusters can be selectively biotinylated, and then isolated using streptavidin. The biotin tag can be subsequently removed using reducing agents (e.g. glutathione).

Targeted and Reversible Biotinylation with Biotin-SS-Tyramide

Tyramine derivatives such as Biotin-Tyramide are converted to radicals by horseradish peroxidase (HRP) in the presence of H₂O₂. These highly reactive species preferentially react with tyrosine residues of proteins, a property that is employed in many applications such as protein labeling, in situ hybridization, electron microscopy, ELISA, phage display and others. Biotin-Tyramide is a valuable tool for signal amplification and can significantly improve signal-to-noise ratios.

Biotinylation of proteins by reaction with the Tyramide moiety of Biotin-SS-Tyramide

Biotin-SS-Tyramide is a variant of Biotin-Tyramide that can be cleaved using reducing agents such as glutathione. One possible application of Biotin-SS-Tyramide is the targeted biotinylation of proteins and protein clusters on cell surfaces using HRP-labeled antibodies. Labeled proteins are then purified by using streptavidin and released by reductive cleavage of the linker disulfide bond.

References:

New Insights into the DT40 B Cell Receptor Cluster Using a Proteomic Proximity Labeling Assay; X.-W. Li, J. S. Rees, P. Xue, H. Zhang, S. W. Hamaia, B. Sanderson, P. E. Funk, R. W. Farndale, K. S. Lilley, S. Perrett and A. P. Jackson; Journal of Biological Chemistry 2014; 289: 14434-14447. doi:10.1074/jbc.M113.529578
A Catalysis-Based Selection for Peroxidase Antibodies with Increased Activity; J. Yin, J. H. Mills and P. G. Schultz; Journal of the American Chemical Society 2004; 126: 3006-3007. doi:10.1021/ja039198o