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.

• Target protein (1 mg/mL, 100 µg per reaction)

• Poly-glycine label (5 mM, 5 µl per reaction, <10 kDa)

• 37°C incubator

• Microcentrifuge

• Final formulation buffer (e.g. PBS or TBS)

SrtA, sortase A, sortagging, S aureus

Each kit provides sufficient materials to label 3x 100 µg recombinant antibody/protein.

COMPONENTS:

See kit data sheet for detailed protocol.

Store cell line in liquid nitrogen immediately upon receipt. Store other components as described below. Stable for at least 6 months from date of receipt when stored as directed.

Great for the site-specific labeling of recombinant antibodies and proteins.

Popp M. Methods Mol Biol. 2015; 1266:185-98.

Avoid freeze/thaw cycles.

Sortase

Kit