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;
Sortase A
Sortase A
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Regular price
$340 USD
Regular price
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$340 USD
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per
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Product Details
Product Details
Product Specification
| Species | Staphylococcus aureus |
| Synonyms | LPXTG specific sortase A, sortase, Srt A |
| Expression System | E.coli |
| Molecular Weight | 25 kDa (Reducing) |
| Purity | ＞98% by SDS-PAGE |
| Tag | No Tag |
| Physical Appearance | Liquid |
| Storage Buffer | 50mM Tris-HCl, 100mM NaCl, 2.5mM KCl, pH 8.0, 0.04% Tween 20, 20%甘油 |
| Stability & Storage | Store at -25 ~ -15℃ for 1 years，Avoid freeze / thaw cycle |
| Reference | 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. |
Background
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).
Components
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
Protocol
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.
Guidelines
Avoid freeze / thaw cycle
This product is for research use only.
Picture
Picture
Bioactivity
SDS-PAGE
2μg (R: reducing condition, N: non-reducing condition).
SEC-HPLC
98.4%
RP-HPLC
99.6%
