Sortases catalyze 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. Sortase variants are available that may exhibit enhanced activity with the substrate of interest. This kit provides wildtype Sortase and four different mutants to evaluate which sortase is best for the chosen application.
Target protein (1 mg/mL, 10 μg per reaction) Poly-glycine label (5 mM, 1 μl per reaction, <10 kDa) 37°C incubator Microcentrifuge Electrophoresis sample buffer (for SDS-PAGE) or 1 M EDTA solution (for mass spectroscopy)
Format
References
1. Popp M. Methods Mol Biol. 2015; 1266:185-98. 2. Chen L., et al., 2016 Scientific Reports 6: 31899
Storage/Stability
At least 6 months from date of receipt, when stored as directed.
Instructions for Use
See assay protocol for detailed instructions.
Shipping Temperature
-80°C
Warnings
Avoid multiple freeze/thaw cycles.
Disclaimers and Limitations
BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.
End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.