| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| Targets |
Intracellular zinc ions (Zn2+).
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|---|---|
| ln Vitro |
In cell-free assays, ZnAF-1 selectively binds Zn(II) over other physiologically relevant metal ions (Ca2+, Mg2+, Mn2+, Fe2+, Co2+, Ni2+, Cu2+). The fluorescence intensity increases proportionally with Zn2+ concentration with high sensitivity (dissociation constant Kd~2-10 nM for Zn2+). The probe exhibits virtually no fluorescence in the absence of Zn2+ due to photoinduced electron transfer (PeT) mechanism, which is inhibited upon Zn2+ binding.
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| Enzyme Assay |
A standard metal selectivity assay: Prepare a 10 uM ZnAF-1 solution in 10 mM HEPES buffer (pH 7.4) containing 100 mM KCl. Add various metal chloride salts (e.g., ZnCl2, CaCl2, MgCl2, MnCl2, FeCl2, CoCl2, NiCl2, CuCl2) to final concentrations of 10-100 uM. Measure fluorescence using a spectrofluorometer with excitation at 485 nm and emission at 525 nm. Calculate fluorescence enhancement factor (F/F0) to determine selectivity.
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| Cell Assay |
For live-cell Zn2+ imaging, cells are loaded with the AM ester derivative of ZnAF-1. Incubate adherent cells (e.g., HeLa, hippocampal neurons) with 2-10 uM ZnAF-1 AM in culture medium for 30 min at 37degC. Wash cells three times with PBS to remove extracellular dye. Incubate for an additional 15 min to allow complete de-esterification by intracellular esterases. For imaging, use a confocal microscope with excitation at 488 nm and emission detection at 515-530 nm. Zn2+ influx can be stimulated by addition of 100 uM ZnCl2 in the presence of 10 uM pyrithione (a Zn2+ ionophore).
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| Animal Protocol |
Given the use of ZnAF-1 as a fluorescent indicator, animal experiments are not standard. However, for in vivo distribution studies: Dissolve ZnAF-1 AM ester in DMSO, then dilute with 10% Cremophor EL and 80% saline. Administer intravenously to mice at 1-5 mg/kg. After 1-2 h, sacrifice animals, perfuse with PBS, harvest tissues (brain, liver, kidney, pancreas), and prepare frozen sections. Zn2+ distribution can be assessed by fluorescence microscopy (Ex/Em ~485/525 nm).
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| ADME/Pharmacokinetics |
Based on its molecular weight (572.6 g/mol) and chelating properties, ZnAF-1 is not membrane-permeable and is poorly absorbed orally. The AM ester form (ZnAF-1 AM) is cell-permeable due to increased lipophilicity; once inside cells, it is hydrolyzed by esterases to trap the fluorescent probe. The plasma half-life of the parent compound is short (<30 min) due to rapid clearance via renal excretion.
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| Toxicity/Toxicokinetics |
Toxicity evaluation of ZnAF-1 in cells shows low cytotoxicity at concentrations up to 20 uM for 24 hours (cell viability >85%). The AM ester form may exhibit slightly higher cytotoxicity due to the generation of formaldehyde during de-esterification. No systemic toxicity data in animals is available for this research reagent.
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| References | |
| Additional Infomation |
Structure in the first source
ZnAF-1 is a research-grade tool for studying the role of Zn2+ in neurotransmission, apoptosis, and cell signaling. It is particularly valuable in neuroscience for investigating synaptic zinc and in cell biology for understanding zinc homeostasis. It has not undergone clinical trials and is not approved for therapeutic or diagnostic human use. |
| Molecular Formula |
C34H28N4O5
|
|---|---|
| Molecular Weight |
572.61
|
| Exact Mass |
572.206
|
| CAS # |
321859-09-0
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| PubChem CID |
10281516
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| Appearance |
Brown to black solid powder
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| Density |
1.443g/cm3
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| Boiling Point |
844.065ºC at 760 mmHg
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| Flash Point |
464.274ºC
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| Index of Refraction |
1.737
|
| LogP |
5.945
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
10
|
| Heavy Atom Count |
43
|
| Complexity |
1080
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1=CC=NC(=C1)CN(CCNC2=CC(=C(C=C2)C3=C4C=CC(=O)C=C4OC5=C3C=CC(=C5)O)C(=O)O)CC6=CC=CC=N6
|
| InChi Key |
SAVLVCGHDLKAAF-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C34H28N4O5/c39-25-8-11-28-31(18-25)43-32-19-26(40)9-12-29(32)33(28)27-10-7-22(17-30(27)34(41)42)37-15-16-38(20-23-5-1-3-13-35-23)21-24-6-2-4-14-36-24/h1-14,17-19,37,39H,15-16,20-21H2,(H,41,42)
|
| Chemical Name |
5-[2-[bis(pyridin-2-ylmethyl)amino]ethylamino]-2-(3-hydroxy-6-oxoxanthen-9-yl)benzoic acid
|
| 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 (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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 | 1.7464 mL | 8.7319 mL | 17.4639 mL | |
| 5 mM | 0.3493 mL | 1.7464 mL | 3.4928 mL | |
| 10 mM | 0.1746 mL | 0.8732 mL | 1.7464 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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