Sortase A, S. aureus

The product can be diluted with sterile pure water to 0.5 μg/μL as the working solution.

Preparation of polypeptide reaction system (30 μL system)
1. Test group: 5 μL Srt3 (1 mM), 5 μL Srt4 (1 mM), 5 μL Sortase A (0.5 μg/μL), 10 μL 3× reaction Buffer, 5 μL ddH2O;
2. Control group: 5 μL Srt3 (1 mM), 5 μL Srt4 (1 mM), 20 μL ddH₂O
Prepare the corresponding control group and test group in turn according to the above table. After the preparation, take the test group reagent, mix it evenly, centrifuge it, and react it at 30°C for 1-3h. After the reaction is completed, perform SDS-PAGE electrophoresis on the control group and the test group simultaneously.

Notes
1.3×Reaction Buffer: 100 mM Tris-HCl 7.5, 50 mM CaCl2, 20 mM DTT
2.Synthetic peptides are used for performance reaction.
The peptide sequence is as follows:
SrtA-3 YGVRLCGREFIRAVIFTCGGSRWRRKGGLPRTGG 34
SrtA-4 GGGGSGGRRSRRSRQDLQTLCCTDGCSMTDLSALC 35
It is recommended to divide the synthesized peptide into small tubes. Sterile purified water can be used to dissolve the peptide. It is recommended to prepare it before use.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.