Bacterial Sortase Substrate III, Abz/DNP |
Catalog No.GC35460
One-Click Copy Product Info
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Bacterial Sortase Substrate III, Abz/DNP is a fluorescent quenching substrate (FRET substrate) used to detect bacterial Sortase activity.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 2857862-12-3
Sample solution is provided at 25 µL, 10mM.
Bacterial Sortase Substrate III, Abz/DNP is a fluorescent quenching substrate (FRET substrate) used to detect bacterial Sortase activity. Bacterial Sortase Substrate III, Abz/DNP consists of a Sortase recognition sequence (LPXTG), an N-terminal fluorescent group Abz, and a C-terminal quencher group DNP, and is commonly used for Sortase A (SrtA) enzyme activity assays and inhibitor screening experiments. After cleavage by Sortase A, Abz is separated from DNP, resulting in the recovery of fluorescence. Cleavage of this substrate can be monitored at Ex/Em=320nm/420nm[1].
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.
Sortase A Activity Assay[1-2]
This plan is based on literature, please modify it to meet your specific needs.
1. Preparation of substrate working solution
(1) Prepare storage solution: Dissolve Bacterial Sortase Substrate III, Abz/DNP in DMSO at concentration of 10mM.
Note: Unused storage solution should be aliquoted and stored in the dark at -20°C or -80°C to avoid repeated freezing and thawing.
(2) Prepare working solution: Dilute the stock solution with reaction buffer (50mM Tris-HCl, 150mM NaCl; pH=7.5).
Note: Prepare the working solution fresh and store briefly at 4°C or on ice.
2. Prepare a 96-well plate, including:
Experimental wells: a mixture of Bacterial Sortase Substrate III, Abz/DNP substrate and Sortase A, concentrations of 10μM and 5-15μM, respectively.
Sample blank wells: Bacterial Sortase Substrate III, Abz/DNP substrate only, concentration 10μM, add reaction buffer to make up the volume.
Substrate blank wells: Sortase A only, concentration 5-15μM, add reaction buffer to make up the volume.
3. Incubate the 96-well plate at 37℃ for 10-60 min, and detect the fluorescence intensity at 320nm under 420nm excitation using a microplate reader.
Precautions:
1) Fluorescent substrate has quenching problems. Please avoid light as much as possible to slow down fluorescence quenching.
2) For your safety and health, please wear a lab coat and disposable gloves.
References:
[1] Gaspar AH, Marraffini LA, Glass EM, Debord KL, Ton-That H, Schneewind O. Bacillus anthracis sortase A (SrtA) anchors LPXTG motif-containing surface proteins to the cell wall envelope. J Bacteriol. 2005;187(13):4646-4655.
[2] Chen Y, Li W, Wang L, Wang B, Suo J. Novel inhibition of Staphylococcus aureus sortase A by plantamajoside: implications for controlling multidrug-resistant infections. Appl Environ Microbiol. 2025;91(1):e0180424.
| Cas No. | 2857862-12-3 | SDF | |
| Formula | C41H57N11O14 | M.Wt | 928 |
| Solubility | Soluble in DMSO | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.0776 mL | 5.3879 mL | 10.7759 mL |
| 5 mM | 215.5 μL | 1.0776 mL | 2.1552 mL |
| 10 mM | 107.8 μL | 538.8 μL | 1.0776 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >99.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 5 reference(s) in Google Scholar.)