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| Targets |
ZnAF-1F targets zinc ions (Zn²⁺) in biological systems by acting as a fluorescent probe. The compound has a high affinity for Zn²⁺ with a dissociation constant (Kd) of 2.2 nM. The mechanism of detection involves photoinduced electron transfer (PET), where the fluorescence of the fluorophore is quenched in the absence of Zn²⁺ and restored upon binding. The interaction with Zn²⁺ ions leads to a significant increase in fluorescence intensity, allowing for precise detection of zinc concentrations in biological samples. The compound's fluorescence is stable under physiological conditions. The presence of electron-withdrawing fluorine atoms shifts the pKa to 4.9, ensuring the probe remains responsive in neutral and slightly acidic conditions. The compound is used to study zinc ion dynamics in chemical reactions and coordination chemistry.
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| ln Vitro |
ZnAF-1F (1 μM) in 100 mM HEPES buffer has a Kon value of 3.5*106 M-1s-1 and a Koff value of 7.7*10-3 s-1 [1]. Zinc sensor ZnAF-1F functions in neutral and mildly acidic environments [2].
In vitro, ZnAF-1F is used as a fluorescent probe for the detection and quantification of zinc ions in biological samples. The compound exhibits high affinity for Zn²⁺ with a Kd of 2.2 nM. Upon binding to Zn²⁺, the fluorescence intensity increases significantly, allowing for sensitive detection of zinc concentrations. The probe's excitation wavelength is 489 nm and emission wavelength is 514 nm. The fluorescence is stable under physiological conditions, with a pKa of 4.9. ZnAF-1F can be used to monitor zinc ion concentrations and distribution within living cells. The compound is used in studies of zinc homeostasis, signaling, and enzyme activity. Its high affinity and sensitivity make it suitable for quantifying zinc ions in complex mixtures. |
| ln Vivo |
ZnAF-1F is not used as a therapeutic agent but rather as a research tool for detecting and studying zinc ions in biological systems. The compound is used in cellular imaging to monitor zinc ion concentrations and distribution within living cells. It assists in understanding the role of zinc in cellular processes such as signaling and enzyme activity. The compound is used in pathophysiological investigations to study the involvement of zinc ions in diseases such as neurodegeneration and cancer. It aids in the design of zinc-based therapeutic agents and diagnostic tools. The compound is also applied in creating zinc-specific sensors for environmental monitoring and clinical diagnostics. ZnAF-1F is used in research to investigate zinc's role in neuronal signaling pathways. The compound is not approved for clinical use.
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| Enzyme Assay |
In vitro experiments with ZnAF-1F typically involve preparing a stock solution in DMSO and diluting it in appropriate buffers for fluorescence measurements. The compound is used at concentrations ranging from 0.1-10 μM depending on the application. For zinc detection, the probe is added to samples containing Zn²⁺, and fluorescence is measured at excitation 489 nm and emission 514 nm. A standard curve is prepared using known concentrations of Zn²⁺ to quantify the amount of zinc in unknown samples. The fluorescence increase upon Zn²⁺ binding is proportional to the zinc concentration. The compound's high affinity (Kd 2.2 nM) allows for detection of low concentrations of zinc. The probe's fluorescence is stable under physiological conditions, with a pKa of 4.9. For cellular uptake studies, the diacetyl derivative (ZnAF-1F DA) may be used to permeate cell membranes.
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| Cell Assay |
In vitro cell-based assays using ZnAF-1F involve loading cells with the probe to monitor intracellular zinc concentrations. The diacetyl derivative (ZnAF-1F DA) is often used because it can permeate cell membranes. Cells are incubated with ZnAF-1F DA at concentrations of 1-10 μM for 30-60 minutes at 37°C. The probe is deacetylated inside cells to the active form, which then binds to intracellular Zn²⁺. Fluorescence is measured using a fluorescence microscope or flow cytometer with excitation at 489 nm and emission at 514 nm. Changes in fluorescence intensity reflect changes in intracellular zinc levels. The probe can be used to monitor zinc fluctuations during cellular processes such as signaling and apoptosis. Cell viability should be assessed to ensure the probe is not toxic at the concentrations used. The compound's fluorescence is stable under physiological conditions.
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| Animal Protocol |
In vivo animal experiments with ZnAF-1F are limited as the compound is primarily used as a fluorescent probe for in vitro and cellular studies. The compound may be used in ex vivo tissue imaging or in vivo imaging applications where zinc detection is required. For in vivo applications, the probe would need to be formulated for appropriate administration routes. The diacetyl derivative (ZnAF-1F DA) may be used for improved cell permeability. The compound's fluorescence properties allow for imaging of zinc distribution in tissues. However, specific in vivo protocols for this compound are not widely reported in the available literature. The compound is intended for research use only and is not approved for clinical applications.
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| ADME/Pharmacokinetics |
ZnAF-1F has a molecular weight of 608.59 g/mol and the formula C34H26F2N4O5. The compound has a dissociation constant (Kd) of 2.2 nM for Zn²⁺. The excitation wavelength is 489 nm and the emission wavelength is 514 nm. The compound operates through photoinduced electron transfer (PET), with fluorescence increasing upon Zn²⁺ binding. The pKa is shifted to 4.9 due to electron-withdrawing fluorine atoms. For long-term storage, the compound is stored at -20°C. In solvent, it is stored at -80°C. The compound is soluble in DMSO. The diacetyl derivative (ZnAF-1F DA) is used for cellular uptake studies. The compound is used in various fields including chemistry, biology, medicine, and industry.
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| Toxicity/Toxicokinetics |
The toxicity of ZnAF-1F has not been extensively characterized as the compound is primarily used as a fluorescent probe for research applications. The compound is intended for research use only and is not for human use. Standard laboratory safety precautions should be followed when handling the compound, including the use of gloves and eye protection. In cell culture applications, the compound is used at concentrations that are generally compatible with cell viability (1-10 μM). Higher concentrations may affect cell viability or cause non-specific fluorescence. The compound is not classified as a highly toxic substance but should be handled with appropriate care. Safety data sheets recommend standard handling procedures for research chemicals. The compound's toxicity in vivo has not been extensively studied.
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| References | |
| Additional Infomation |
ZnAF-1F (CAS 443302-08-7) is a highly efficient fluorophore used as a fluorescent probe for zinc ions (Zn²⁺) in cellular applications. It has a dissociation constant (Kd) of 2.2 nM. The compound has the molecular formula C34H26F2N4O5 and a molecular weight of 608.59 g/mol. ZnAF-1F exhibits excitation at 489 nm and emission at 514 nm. The compound operates through photoinduced electron transfer (PET), with fluorescence increasing upon Zn²⁺ binding. The pKa is shifted to 4.9 due to electron-withdrawing fluorine atoms. The compound is used for intracellular zinc measurement and zinc homeostasis studies. It is applied in chemistry for fluorescent probing and quantification of zinc ions, in biology for cellular imaging and zinc homeostasis studies, in medicine for pathophysiological investigations and drug development, and in industry for sensor development. The diacetyl derivative (ZnAF-1F DA) is used for cellular uptake studies. The compound is stored at -20°C.
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| Molecular Formula |
C34H26F2N4O5
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| Molecular Weight |
608.59100
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| Exact Mass |
608.187
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| CAS # |
443302-08-7
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| PubChem CID |
16218749
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| Appearance |
Orange to red solid powder
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| Density |
1.49g/cm3
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| Boiling Point |
822.1ºC at 760 mmHg
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| Flash Point |
451ºC
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| Index of Refraction |
1.715
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| LogP |
6.224
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
11
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
45
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| Complexity |
1170
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
PUDBCJSXTISPOO-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C34H26F2N4O5/c35-27-14-25-31(16-29(27)41)45-32-17-30(42)28(36)15-26(32)33(25)23-8-7-20(13-24(23)34(43)44)39-11-12-40(18-21-5-1-3-9-37-21)19-22-6-2-4-10-38-22/h1-10,13-17,39,41H,11-12,18-19H2,(H,43,44)
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| Chemical Name |
5-[2-[bis(pyridin-2-ylmethyl)amino]ethylamino]-2-(2,7-difluoro-3-hydroxy-6-oxoxanthen-9-yl)benzoic acid
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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) |
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 | 1.6431 mL | 8.2157 mL | 16.4314 mL | |
| 5 mM | 0.3286 mL | 1.6431 mL | 3.2863 mL | |
| 10 mM | 0.1643 mL | 0.8216 mL | 1.6431 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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