Description
Sortases catalize a transpeptidation reaction, where the enzyme cleaves the amide bond between the threonine and glycine of the sortase recognition sequence (LPXTG for S. aureus Sortase A), generating a thioacyl intermediate. Subsequently, this intermediate is resolved by the N-terminus of an oligoglycine nucleophile, creating a new peptide bond that links the substrate to the incoming nucleophile. The Sortagging Kit takes advantage of this reaction to label virtually any recombinant antibody or protein containing the sortase recognition sequence. Labeling efficiency of greater than 90% is achieved by taking advantage of the highly active Sortase 5Δ Pentamutant. Simply combine your protein of interest with Sortase 5Δ and a polyglycine label, and incubate for 1 hour at 37°C. Purification columns are included to remove Sortase 5Δ and excess label. The kit also includes Ubiquitin-LPETGH6 and GGG-Clover as positive controls