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One-Step Fluorescent Protein Labeling by Tubulin Tyrosine Ligase

Methods Mol Biol. 2019; 
Schumacher D, Leonhardt H, Hackenberger CPR, Helma J.
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Molecular Biology Reagents 1. Expression of pET plasmid (Genscript) coding for the FN3CD20 protein and a hexahistidine-tag at the C-terminus (see Note 1). Get A Quote

摘要

Tub-tag labeling, a novel chemoenzymatic protein functionalization method, facilitates one-step fluorescent labeling of functional biomolecules. The enzyme tubulin tyrosine ligase incorporates coumarin-amino acids to the terminal carboxylic acid of proteins containing a short peptidic recognition sequence called Tub-tag. Here we describe the one-step Tub-tag protein modification protocol in detail and explain its utilization to generate fluorescently labeled proteins for advanced applications in imaging and diagnostics.

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