| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| Targets |
The target of FP-Biotin is the catalytic serine residue found in the active site of serine hydrolases. This is a large enzyme superfamily that includes carboxylesterases, cholinesterases, lipases, and peptidases. FP-Biotin irreversibly binds to this nucleophilic serine, covalently labeling the enzyme in an activity-dependent manner.
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|---|---|
| ln Vitro |
Despite having a large biotin group, FP-biotin displays high reactivity with human AChE and BChE. For the suggested goal of detecting reactive proteins associated with organophosphorus toxicants, FP-biotin is appropriate[1].
As a chemical probe, FP-Biotin is validated for labeling several key serine hydrolases, including Fatty Acid Amide Hydrolase (FAAH). In vitro, it reacts rapidly and with high sensitivity with crude tissue extracts, labeling numerous serine hydrolases. These labeled proteins can be visualized on a gel or enriched for mass spectrometry identification. |
| ln Vivo |
There are no in vivo activity data for FP-Biotin as it is a tool for ex vivo proteomic analysis. However, it has proven invaluable in identifying novel biomarkers of exposure to organophosphorus toxicants in vivo by measuring the residual activity of serine hydrolases, such as FAAH, ABHD6, and MAG-lipase, in plasma samples.
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| Enzyme Assay |
A standard non-cellular protocol for FP-Biotin is a competitive ABPP assay. Tissue lysates (e.g., mouse brain) are pre-incubated with a serine hydrolase inhibitor of interest (or DMSO) for 30 min at room temperature. Then, FP-Biotin (e.g., 5 uM) is added and incubated for 30 min. The reaction is stopped with SDS loading buffer, and samples are resolved by SDS-PAGE.
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| Cell Assay |
Cell-based protocols are not applicable for FP-Biotin, as it is used to analyze cell lysates. For cell culture, cells are harvested, washed with PBS, and lysed via sonication in a suitable buffer (e.g., 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% Triton X-100). The lysate is centrifuged, and the supernatant (whole proteome) is used for the labeling reaction described in the non-cellular section.
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| Animal Protocol |
FP-Biotin is used in ex vivo assays from animal experiments. Mice are treated with a test compound (e.g., a FAAH inhibitor). After the treatment period, the animals are sacrificed, and tissues (e.g., brain, liver) are harvested. Tissues are homogenized, and lysates are prepared. The lysates are then labeled with FP-Biotin as described in the non-cellular protocol.
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| ADME/Pharmacokinetics |
As a chemical probe, FP-Biotin itself does not have classical pharmacokinetic parameters, as it is not a therapeutic agent. It is a reactive compound used for ex vivo analysis. Its stability and reactivity in a given buffer are the relevant properties. It is stable when stored at -20degC as a powder, but it is reactive in solution.
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| Toxicity/Toxicokinetics |
The toxicological profile of FP-Biotin is not relevant for its use, as it is used at low concentrations in protein extracts. The compound is a potent organophosphorus compound and should be handled with care as a potential toxicant. Standard laboratory safety procedures for handling reactive chemical probes should be followed.
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| References |
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| Additional Infomation |
FP-Biotin is arguably one of the most important chemical tools for the field of chemical biology. It was the prototype that enabled Activity-Based Protein Profiling (ABPP), transforming it from a gel-based visualization technique to a powerful mass spectrometry-driven discovery platform. The biotin tag allows for the efficient purification of labeled peptides through binding to immobilized avidin beads.
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| Molecular Formula |
C27H50FN4O5PS
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|---|---|
| Molecular Weight |
592.75
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| Exact Mass |
592.322
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| CAS # |
259270-28-5
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| PubChem CID |
24762158
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| Appearance |
White to light yellow solid powder
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| Density |
1.125g/cm3
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| Boiling Point |
851.3ºC at 760 mmHg
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| Flash Point |
468.6ºC
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| Vapour Pressure |
2.82E-29mmHg at 25°C
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| Index of Refraction |
1.497
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| LogP |
7.084
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
23
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| Heavy Atom Count |
39
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| Complexity |
793
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCOP(=O)(CCCCCCCCCC(=O)NCCCCCNC(=O)CCCCC1C2C(CS1)NC(=O)N2)F
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| InChi Key |
ZIFSFLGSUTZFCP-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C27H50FN4O5PS/c1-2-37-38(28,36)20-14-7-5-3-4-6-9-16-24(33)29-18-12-8-13-19-30-25(34)17-11-10-15-23-26-22(21-39-23)31-27(35)32-26/h22-23,26H,2-21H2,1H3,(H,29,33)(H,30,34)(H2,31,32,35)
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| Chemical Name |
10-[ethoxy(fluoro)phosphoryl]-N-[5-[5-(2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl)pentanoylamino]pentyl]decanamide
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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 (e.g. under nitrogen), 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: 25 mg/mL (42.18 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.22 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 (4.22 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.6871 mL | 8.4353 mL | 16.8705 mL | |
| 5 mM | 0.3374 mL | 1.6871 mL | 3.3741 mL | |
| 10 mM | 0.1687 mL | 0.8435 mL | 1.6871 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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