| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 50mg |
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| Targets |
HADA hydrochloride targets the peptidoglycan (PG) biosynthesis machinery of bacteria. Peptidoglycan is a polymer consisting of sugars and amino acids that forms a mesh-like layer outside the plasma membrane of most bacteria, providing structural integrity and protection. HADA is a fluorescent D-amino acid that is incorporated into the peptidoglycan at sites of active synthesis by the bacterial transpeptidase and transglycosylase enzymes. Because D-amino acids are not typically used by mammalian cells, HADA specifically labels bacterial cells without affecting eukaryotic cells. The compound's ability to label peptidoglycan makes it a valuable tool for studying bacterial growth, cell division, and antibiotic susceptibility.
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| ln Vitro |
FDAA labeling for organisms that develop quickly (like E. Coli) only takes 30 seconds. HADA hydrochloride offers the most reliable and consistent PG labeling for the majority of bacterial species and typically does not require significant adjustment, despite being less photostable and darker than FDL or TDL [1].
In vitro, HADA hydrochloride is used for the fluorescent labeling of peptidoglycans in live bacteria. The compound is efficiently incorporated into the peptidoglycan at sites of biosynthesis, allowing the visualization of bacterial cell walls by fluorescence microscopy. HADA labeling results in strong peripheral and septal staining of diverse bacterial cell populations, including both Gram-positive and Gram-negative bacteria. The labeling is rapid, with incorporation observed within 30 seconds for rapidly growing species such as E. coli. The compound does not affect bacterial growth rates, making it suitable for time-lapse imaging of bacterial growth and division. HADA can also be used in combination with other fluorescent probes for multiplexed imaging. |
| ln Vivo |
In vivo, HADA hydrochloride is used for labeling bacteria in living organisms, such as in animal models of infection or in the gut microbiome. The compound can be administered to animals to label bacteria in situ, allowing the visualization of bacterial populations and their spatial distribution in tissues. HADA labeling has been used to study the dynamics of bacterial colonization, biofilm formation, and the effects of antibiotics on bacterial growth in vivo. The compound's specificity for bacterial peptidoglycan makes it a valuable tool for studying host-bacteria interactions and for tracking bacterial pathogens in infection models. The fluorescence of HADA allows for real-time imaging of bacterial growth and division in living animals.
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| Enzyme Assay |
In vitro binding/incorporation assays for HADA hydrochloride are performed by incubating bacteria with the fluorescent probe. Bacteria (e.g., E. coli, Bacillus subtilis) are grown to mid-log phase and incubated with HADA at concentrations ranging from 1 to 100 μM for 10-60 minutes. After washing to remove unbound probe, bacteria are fixed and examined by fluorescence microscopy. The incorporation of HADA into peptidoglycan is visualized as fluorescent labeling of the cell wall, with characteristic patterns depending on the bacterial species and growth phase. The labeling efficiency can be quantified by flow cytometry or by measuring fluorescence intensity. The assay includes controls (bacteria incubated without HADA or with non-fluorescent D-amino acids).
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| Cell Assay |
In vitro cell-based assays for HADA hydrochloride are performed using bacterial cultures grown in liquid media or on agar plates. Bacteria are incubated with HADA (typically 10-50 μM) for 10-60 minutes, depending on the species and growth rate. After labeling, bacteria are washed and examined by fluorescence microscopy. HADA labeling allows the visualization of bacterial morphology, cell division, and the effects of antibiotics on cell wall synthesis. Time-lapse microscopy can be used to track bacterial growth and division in real-time. The compound can be used in combination with other fluorescent dyes (e.g., membrane dyes, nucleic acid stains) for multiplexed imaging. The labeling efficiency and specificity are assessed by comparing fluorescence intensity with controls.
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| Animal Protocol |
In vivo animal studies with HADA hydrochloride are conducted in mouse models of bacterial infection. Mice are infected with a bacterial pathogen (e.g., E. coli, Staphylococcus aureus) and then administered HADA via intravenous, intraperitoneal, or oral routes. The compound is allowed to circulate and label bacteria in vivo. After a suitable labeling period, tissues are collected, and bacterial labeling is visualized by fluorescence microscopy or flow cytometry. The distribution of bacteria in tissues and the effects of antibiotics on bacterial growth can be assessed. The compound's specificity for bacterial peptidoglycan allows the selective labeling of bacteria in the presence of host cells.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of HADA hydrochloride have not been extensively characterized. As a small fluorescent D-amino acid (molecular weight 328.71), the compound is expected to have moderate aqueous solubility and membrane permeability. HADA is soluble in DMSO at 125 mg/mL and can be diluted in aqueous buffers for biological applications. The compound's stability in biological fluids and its tissue distribution would need to be determined for specific applications. For in vivo labeling, the compound is typically administered in saline or other suitable vehicles. The pharmacokinetics of HADA would depend on factors such as the route of administration, dose, and the presence of bacterial targets for the probe to bind to.
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| Toxicity/Toxicokinetics |
HADA hydrochloride is generally considered to have low toxicity, as it is a fluorescent D-amino acid that specifically labels bacteria without affecting eukaryotic cells. The compound does not affect bacterial growth rates at labeling concentrations, indicating minimal interference with bacterial physiology. No specific toxicological studies have been reported in the literature. As a research chemical, standard safety precautions should be followed when handling HADA hydrochloride, including the use of personal protective equipment. The compound should be stored at -20°C and protected from light. HADA hydrochloride is for research use only and not for human or veterinary applications.
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| References | |
| Additional Infomation |
HADA hydrochloride is a member of the family of fluorescent D-amino acids (FDAAs) that are used for the labeling of peptidoglycan in bacteria. These probes have revolutionized the study of bacterial cell wall dynamics, allowing researchers to visualize bacterial growth, division, and the effects of antibiotics in real-time at the single-cell level. HADA is one of several FDAAs with different fluorescence colors (e.g., NADA, green; HADA, blue; TADA, red), enabling multiplexed imaging of bacterial populations. The probes are widely used in microbiology research to study bacterial physiology, host-pathogen interactions, and antibiotic mechanisms. HADA hydrochloride is available from chemical suppliers for research purposes only.
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| Molecular Formula |
C13H13CLN2O6
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| Molecular Weight |
328.705122709274
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| Exact Mass |
328.046
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| CAS # |
2253733-10-5
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| Related CAS # |
1417905-41-9;2253733-10-5 (HCl);
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| PubChem CID |
137919867
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
22
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| Complexity |
489
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| Defined Atom Stereocenter Count |
1
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| SMILES |
Cl.O1C(C(C(NC[C@H](C(=O)O)N)=O)=CC2C=CC(=CC1=2)O)=O
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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 : 125 mg/mL (380.27 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.08 mg/mL (6.33 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 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: ≥ 2.08 mg/mL (6.33 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 20.8 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. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (6.33 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.0422 mL | 15.2110 mL | 30.4220 mL | |
| 5 mM | 0.6084 mL | 3.0422 mL | 6.0844 mL | |
| 10 mM | 0.3042 mL | 1.5211 mL | 3.0422 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
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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