| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
5-FAM-Alkyne is designed to target and detect alkaline phosphatase (ALP) activity. The compound itself is not an enzyme inhibitor but rather a fluorescent substrate or probe. Upon dephosphorylation by ALP, the probe releases a fluorescent signal that can be quantitatively measured. The alkyne functionality allows for covalent attachment to azide-modified biomolecules via click chemistry, expanding its utility for labeling and detection applications in complex biological systems.
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| ln Vitro |
A single piece carrying FAM fluorophore can signal fluorescence strongly and diffuse into solution with ease[1].
In vitro, 5-FAM-Alkyne functions as a fluorescent probe for detecting ALP activity. The compound exhibits excellent photophysical properties with an absorption maximum at 490 nm and emission maximum at 513 nm, and a molar extinction coefficient of 80.0 L mmol⁻¹ cm⁻¹. The probe is highly sensitive and selective for ALP, allowing for real-time monitoring of enzyme activity in solution. Upon dephosphorylation, the fluorophore is released and generates a strong fluorescence signal that correlates with ALP concentration. |
| ln Vivo |
In vivo applications of 5-FAM-Alkyne are limited due to its primary use as an in vitro labeling and detection reagent. The compound's fluorescence properties make it potentially suitable for imaging applications, but its utility in living organisms has not been extensively characterized. The probe may be used in ex vivo tissue staining or in cell-based fluorescence imaging studies. However, the compound is primarily designed for biochemical and molecular biology applications rather than in vivo pharmacology.
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| Enzyme Assay |
The in vitro enzyme assay for 5-FAM-Alkyne involves incubating the probe with ALP enzyme in appropriate buffer (typically Tris-HCl or glycine buffer, pH 8-10) at 37°C. The reaction is monitored in real-time by measuring fluorescence at excitation 490 nm and emission 513 nm using a fluorescence spectrophotometer or plate reader. ALP activity is quantified by the rate of fluorescence increase over time. Controls include reactions without enzyme and with known ALP inhibitors to verify specificity.
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| Cell Assay |
For in vitro cell-based assays, cells are cultured and lysed, and the cell lysate is incubated with 5-FAM-Alkyne probe in assay buffer. Fluorescence is measured to determine ALP activity in the cell lysate. Alternatively, live cells can be incubated with the probe and fluorescence is measured using flow cytometry or fluorescence microscopy. The probe can also be used for immunolabeling applications where ALP-conjugated secondary antibodies are detected.
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| Animal Protocol |
In vivo animal studies for 5-FAM-Alkyne are not commonly performed, as the compound is primarily a research reagent for in vitro applications. If used in vivo, the probe would be administered via intravenous injection and fluorescence imaging would be performed to detect ALP activity in tissues. However, such applications are not well-established and the compound's pharmacokinetics and biodistribution have not been characterized.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of 5-FAM-Alkyne have not been extensively characterized, as the compound is used primarily as a research reagent rather than a drug candidate. The probe is soluble in aqueous buffer at pH >8, as well as in alcohols, DMSO, and DMF. Stability in solution may be limited, and it is recommended to prepare fresh solutions for each experiment. The compound is stable as a solid when stored under recommended conditions.
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| Toxicity/Toxicokinetics |
Toxicology data for 5-FAM-Alkyne are limited. The compound is considered a research chemical and should be handled with standard laboratory safety precautions. As a fluorescent probe, it is not intended for therapeutic use and its toxicity profile has not been systematically evaluated. The compound may be irritating to skin and eyes, and appropriate personal protective equipment should be used when handling.
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| References | |
| Additional Infomation |
5-FAM-Alkyne is a versatile fluorescent probe for ALP detection and bioconjugation applications. The compound's green fluorescence, high sensitivity, and alkyne functionality make it a valuable tool for enzyme activity assays, protein labeling, and click chemistry applications. It is widely used in biochemical research, diagnostic assay development, and molecular biology. The probe is intended for research use only and is not approved for clinical diagnostic or therapeutic applications.
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| Molecular Formula |
C24H15NO6
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|---|---|
| Molecular Weight |
413.38
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| Exact Mass |
413.089
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| CAS # |
510758-19-7
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| PubChem CID |
89857150
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| Appearance |
Yellow to brown solid powder
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| Density |
1.6±0.1 g/cm3
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| Boiling Point |
709.6±60.0 °C at 760 mmHg
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| Melting Point |
313 °C(dec.)
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| Flash Point |
382.9±32.9 °C
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| Vapour Pressure |
0.0±2.3 mmHg at 25°C
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| Index of Refraction |
1.767
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| LogP |
2
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
31
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| Complexity |
762
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C#CCNC(=O)C1=CC2=C(C=C1)C3(C4=C(C=C(C=C4)O)OC5=C3C=CC(=C5)O)OC2=O
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| InChi Key |
UNBAJBIGCBVWJU-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C24H15NO6/c1-2-9-25-22(28)13-3-6-17-16(10-13)23(29)31-24(17)18-7-4-14(26)11-20(18)30-21-12-15(27)5-8-19(21)24/h1,3-8,10-12,26-27H,9H2,(H,25,28)
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| Chemical Name |
3',6'-dihydroxy-3-oxo-N-prop-2-ynylspiro[2-benzofuran-1,9'-xanthene]-5-carboxamide
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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 |
| 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: 50 mg/mL (120.95 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.05 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% 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 25.0 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: ≥ 2.5 mg/mL (6.05 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 25.0 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4191 mL | 12.0954 mL | 24.1908 mL | |
| 5 mM | 0.4838 mL | 2.4191 mL | 4.8382 mL | |
| 10 mM | 0.2419 mL | 1.2095 mL | 2.4191 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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