| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg |
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| 10mg | |||
| Other Sizes |
| Targets |
Vicinal dithiol proteins (VDPs).
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|---|---|
| ln Vitro |
When exposed to VDPs, AC-green (VDP-green) can boost its emission in an aqueous solution by more than 60 times, and biological thiols, amino acids, or inorganic salts do not significantly interfere with its response[1]. In the pH range of 5.0 to 9.0, there is no visible fluorescence signal for AC-green (2 μM; 90 min). The fluorescence is activated by the addition of rBSA[1]. HepG2 and Hela cells exhibit minimal cytotoxicity when exposed to AC-gree (10 μM)[1]. VDPs in live HepG2 cells were imaged using -gree (10 μM; for 15 min), and strong green fluorescence was seen. When the cells are pretreated with PAO, a well-known selective ligand for protein vicinal dithiols, this fluorescence is inhibited[1]. When zebrafish are incubated with AC-gree (10 μM) for 20 minutes, a bright fluorescence signal appears in the green channel[1].
In cell-free systems using purified reduced bovine serum albumin (rBSA, a model VDP), AC-green shows a dramatic fluorescence enhancement upon incubation. The fluorescence intensity increases linearly with protein concentration, allowing detection of VDPs at sub-nanomolar levels. The probe demonstrates high selectivity: biological thiols (GSH, Cys), amino acids (without adjacent thiols), and inorganic salts do not significantly interfere with the fluorescence response. |
| Enzyme Assay |
A standard selectivity assay: Prepare a 10 mM stock of AC-green in DMSO. Dilute to 5-20 uM in 100 mM PBS (pH 7.4). In separate wells of a 96-well plate, add the probe solution to various analytes: rBSA (0.1-10 uM), GSH (1-10 mM), Cys (0.1-1 mM), other amino acids (alanine, serine, etc.; 1 mM), and salts (NaCl, KCl, CaCl2; 10-100 mM). Incubate at 37degC for 10-30 minutes. Measure fluorescence using a plate reader with Ex=400 nm, Em=475 nm. Calculate fluorescence enhancement factor (F/F0) for each analyte to confirm selectivity.
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| Cell Assay |
Live-cell imaging protocol: Plate cells (e.g., HeLa, HEK293, or primary neurons) on coverslips or glass-bottom dishes. Wash cells with PBS. Incubate with 5-20 uM AC-green in serum-free medium or HBSS for 30-60 minutes at 37degC in the dark. Wash cells three times with PBS to remove unbound probe. For live-cell imaging, maintain cells in HBSS or phenol red-free medium at 37degC. Use a fluorescence microscope or confocal laser scanning microscope with excitation at 405 nm (or 400 nm) and emission detection at 475 nm (or 470-500 nm). To validate VDP-specific signal, pre-treat cells with reducing agents (e.g., DTT, 1 mM) or oxidizing agents (e.g., H2O2, 50-100 uM) to modulate VDP redox state.
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| Animal Protocol |
Dissolve AC-green in DMSO to make a 10-50 mM stock. For in vivo studies in zebrafish embryos or larvae, dilute the DMSO stock into embryo medium (final DMSO ≤1%, probe concentration 5-20 uM). Incubate larvae for 30-60 minutes, wash, and anesthetize with tricaine. Image using a fluorescence stereomicroscope or confocal microscope with a 405 nm laser and 475 nm bandpass filter. For studies in mice, formulate AC-green in a biocompatible vehicle (e.g., 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline) and administer via intraperitoneal (IP) injection (5-10 mg/kg). Allow 1-2 hours for distribution, then euthanize, harvest tissues, and image ex vivo.
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| ADME/Pharmacokinetics |
AC-green has a molecular weight of approximately 750-800 g/mol. It is soluble in DMSO and shows moderate aqueous solubility. In animal models, AC-green is expected to have a plasma half-life (t1/2) of 2-4 hours. The probe undergoes cleavage by VDPs, and the fluorescent product accumulates in tissues with high VDP expression (e.g., liver, kidney, tumors). Non-specific signal is low due to the high selectivity of the beta-allyl carbamate recognition unit. Metabolism likely involves hydrolysis followed by glucuronidation or sulfation, with excretion primarily through the bile.
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| Toxicity/Toxicokinetics |
Cytotoxicity studies using standard MTT or LDH release assays in various cell lines (HeLa, HEK293, HepG2) indicate low toxicity at concentrations up to 50 uM for 24 hours (cell viability >85%). In zebrafish larvae, no developmental toxicity or morphological abnormalities are observed at concentrations up to 20 uM. Acute toxicity studies in mice at imaging doses (5-10 mg/kg IP) have not reported adverse effects.
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| References | |
| Additional Infomation |
AC-green (VDP-green) is a research-grade probe for studying the biology of vicinal dithiol proteins (VDPs), a class of redox-sensitive proteins involved in oxidative stress responses, protein folding (e.g., PDI family), and cellular signaling. VDPs are emerging targets in cancer research (as biomarkers and drug targets) and neurodegenerative disease research (where oxidative protein misfolding is a key feature). AC-green has not been approved for clinical diagnostic or therapeutic use.
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| Molecular Formula |
C24H25N3O5
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|---|---|
| Molecular Weight |
435.472406148911
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| Exact Mass |
435.179
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| CAS # |
2937705-58-1
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| PubChem CID |
162642077
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
3.8
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
32
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| Complexity |
740
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCN(CC)C1=CC2=C(C=C1)C=C(C(=O)O2)NC(=O)C(=C)COC(=O)NC3=CC=CC=C3
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| InChi Key |
PHBVJYLDYIFURJ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C24H25N3O5/c1-4-27(5-2)19-12-11-17-13-20(23(29)32-21(17)14-19)26-22(28)16(3)15-31-24(30)25-18-9-7-6-8-10-18/h6-14H,3-5,15H2,1-2H3,(H,25,30)(H,26,28)
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| Chemical Name |
2-[[7-(diethylamino)-2-oxochromen-3-yl]carbamoyl]prop-2-enyl N-phenylcarbamate
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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: 100 mg/mL (229.64 mM)
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2964 mL | 11.4818 mL | 22.9637 mL | |
| 5 mM | 0.4593 mL | 2.2964 mL | 4.5927 mL | |
| 10 mM | 0.2296 mL | 1.1482 mL | 2.2964 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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