25 kDa (Reducing)
Store at -25 ~ -15℃ for 1 years，Avoid freeze / thaw cycle
1. Frankel B A, Kruger R G , Robinson D E ,et al. Staphylococcus aureus sortase transpeptidase SrtA: insight into the kinetic mechanism and evidence fora reverse protonation catalytic mechanism.[J].Biochemistry, 2005, 44(33):11188-11200.
2. Parthasarathy R, Subramanian S, Boder E T. Sortase A as a Novel Molecular "Stapler" for Sequence-Specific Protein Conjugation[J]. Bioconjugate Chemistry, 2007, 18(2):469-476.
Sortase is a family of transpeptidases found in Gram-positive bacteria, which is divided into two subfamilies: Sortase A and Sortase B. Its main function is to catalyze the covalent attachment of cell surface proteins to the cell wall. Among them, Sortase A (SrtA) has been studied more extensively. Sortase A (SrtA) can specifically recognize proteins with a C-terminal conserved amino acid sequence LPXTG and an N-terminal polyglutamine (X is any amino acid except cysteine and tryptophan).
0.5 ug/ul Sortase A in 50mM Tris-HCl, 100mM NaCl, 2.5mM KCl, 0.04% Tween 20，20%甘油, pH 8.0 @ 25°C
3×Reaction Buffer：100mM Tris-HCl、50mM CaCl2、20mM DTT, pH 7.5 @ 25°C
1. Typical polypeptide reaction conditions are as follows:
Components | Volume | Final Concentration |
Polypeptide 1 (1 mM) | 5 µl | 166 uM |
Polypeptide 2 (1 mM) | 5 µl | 166 uM |
Sortase A (0.5 ug/ul) | 5 µl | - |
3X Reaction Buffer | 10 µl | 1X |
ddH2O | Up to 30µl | - |
2. After preparation according to the above table, mixed centrifugation, reaction at 30℃ for 1-3h, and SDS-PAGE electrophoresis was performed after reaction.
Avoid freeze / thaw cycle
This product is for research use only.
Sortase A can specifically identify the peptide sequence LPXTG. Synthetic peptide 1 (YGVRLCGREFIRAVIF-TCGGSRWRRKGGLPRTGG) and peptide 2 (GGGGSGGRRSRRSRQDLQTLCCTDGCSMTDLSALC) to connect, conduct experiments and tests.
Lane 1 Polypeptide 1;
Lane 2 Polypeptide 2;
Lane 3 Negative Control (negative control with no added enzyme only);
Lane 4 UA-Sortase A: 1 ug;
Lane 5 UA-Sortase A: 2.5 ug;
Lane 6 UA-Sortase A: 4 ug;
Lane 7 UA-Sortase A: 5 ug;
2μg (R: reducing condition, N: non-reducing condition).
98.4%
99.6%
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