| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| Other Sizes |
| Targets |
This substrate targets the Gram‑positive bacterial transpeptidase sortase A (SrtA), a membrane‑bound cysteine enzyme that anchors virulence and colonisation‑associated surface proteins to the bacterial cell wall. It serves as a tool for studying sortase‑mediated protein anchoring and for screening sortase inhibitors.
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| ln Vitro |
In FRET‑based assays, the intact substrate shows minimal fluorescence because the Abz fluorophore is quenched by the Dnp group. Upon specific cleavage by SrtA at the threonine‑glycine bond, the quencher is released, resulting in a measurable increase in fluorescence at Ex/Em=320/420 nm. Km ~5.5 mM, kcat ~0.27 s-¹.
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| ln Vivo |
No direct in vivo activity is described, as the substrate is designed for in vitro enzymatic assays. However, understanding SrtA activity in vivo is critical for developing anti‑infective agents, and this substrate can be used to evaluate compound efficacy in cell‑based or ex vivo models of bacterial infection.
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| Enzyme Assay |
A standard SrtA assay protocol: Dilute purified SrtA enzyme into assay buffer (50 mM HEPES, 150 mM NaCl, 5 mM CaCl2, pH 7.5). Add Bacterial Sortase Substrate III, Abz/DNP TFA (final concentration 10‑200 uM) to initiate the reaction. Incubate at 37degC for 30‑60 min. Monitor fluorescence increase at Ex=320 nm/Em=420 nm in real‑time using a microplate reader. Controls without enzyme are included for background subtraction.
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| Cell Assay |
For cellular assays, SrtA‑expressing Staphylococcus aureus cells are cultured in appropriate medium and incubated with the substrate (10‑50 uM) for 1‑4 h. Following incubation, cells are lysed or the supernatant is collected, and fluorescence is measured at Ex/Em=320/420 nm to assess SrtA activity within the bacterial environment.
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| Animal Protocol |
A general animal protocol for infection studies: BALB/c mice (n=6/group) are infected intravenously with S. aureus (1‑5×10⁷ CFU). Test sortase inhibitors are administered orally or intraperitoneally. After 24‑48 h, bacterial loads in organs (kidneys, spleen) are quantified by CFU plating. The substrate is not directly administered but is used for ex vivo activity measurements from isolated bacteria or tissue homogenates.
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| ADME/Pharmacokinetics |
As a peptide substrate, Bacterial Sortase Substrate III, Abz/DNP TFA is not intended for pharmacokinetic studies. It is used as an assay reagent rather than as a therapeutic agent. The compound is typically dissolved in DMSO for stock solutions and diluted in aqueous buffers immediately before use; it should be stored as a lyophilised powder at -20degC protected from light.
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| Toxicity/Toxicokinetics |
The substrate itself is non‑toxic at assay concentrations (≤200 uM). It is intended for research use only and not for human or animal therapeutic applications. The Dnp group is a well‑established quencher in FRET peptide substrates and is not associated with significant toxicity at the concentrations used in enzymatic assays.
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| References |
[1]. Zhang J, et al. Antiinfective therapy with a small molecule inhibitor of Staphylococcus aureus sortase. Proc Natl Acad Sci U S A. 2014 Sep 16;111(37):13517-22.
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| Additional Infomation |
This substrate contains a LPETG sorting signal motif that mimics the natural recognition sequence for SrtA. Cleavage occurs between the threonine and glycine residues, releasing the Dnp‑labelled C‑terminal fragment. The FRET‑based detection enables continuous, real‑time monitoring of transpeptidase activity, making it suitable for high‑throughput screening of sortase inhibitors for anti‑infective drug discovery.
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| Molecular Formula |
C41H57N11O14.XC2HF3O2
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| Related CAS # |
Bacterial Sortase Substrate III, Abz/DNP;2857862-12-3
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| Appearance |
Light yellow to yellow solid powder
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO :~50 mg/mL
H2O :< 0.1 mg/mL |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.25 mg/mL (Infinity mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 1.25 mg/mL (Infinity mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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