| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
Boc-Lys(Ac)-AMC serves as a substrate for histone deacetylases (HDACs), a family of enzymes that remove acetyl groups from lysine residues on histones and other proteins. Following deacetylation by an HDAC, the Boc-Lys-AMC intermediate is cleaved by trypsin, releasing the fluorescent 7-amino-4-methylcoumarin (AMC) moiety. The fluorescence of released AMC can be quantified and used to measure HDAC activity. This makes the compound a valuable tool for studying HDAC function and screening HDAC inhibitors.
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| ln Vitro |
Guidelines (This is our suggested protocol; it should be adjusted based on your unique requirements as it just offers as a guideline). HDAC detection based on tissue culture[2] 1. In 96-well tissue culture plates, add 6 × 104 cells, 100 mL of culture media, and 25 μM Boc-Lys(Ac)-AMC. Culture the cells overnight. 2. For two to three hours, incubate at 37°C and 5% CO2. 3. To stop the deacetylation reaction, add HADC developing solution, which contains cell lysis buffer and trypsin reagent. 4. For fifteen minutes, incubate at room temperature. 5. To measure the fluorescence signal of AMC at Ex/Em = 355 nm/460 nm, use the SpectraMax M5 microplate analyzer.
In vitro, Boc-Lys(Ac)-AMC is used as a substrate to measure HDAC enzymatic activity. The compound is cell-permeable and can be used in cell-based assays to monitor intracellular HDAC activity. Following deacetylation by HDACs and subsequent cleavage by trypsin, the released AMC emits fluorescence at 460 nm upon excitation at 355 nm (Ex/Em = 355 nm/460 nm). The fluorescence signal is directly proportional to HDAC activity, allowing for quantitative analysis. |
| ln Vivo |
Specific in vivo activity data for Boc-Lys(Ac)-AMC are not applicable as the compound is a substrate used in biochemical assays rather than a therapeutic agent. It is not designed for in vivo administration or pharmacological activity. Its primary application is in vitro as a research tool to measure HDAC activity in cell lysates or living cells. The compound is used in research settings to study HDAC function and to evaluate the potency of HDAC inhibitors.
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| Enzyme Assay |
A typical HDAC activity assay using Boc-Lys(Ac)-AMC involves incubating the substrate with an HDAC enzyme source (such as recombinant HDAC protein or cell lysates containing HDAC activity). The reaction is carried out in a suitable HDAC assay buffer at 37°C for a specified time. After incubation, a developer solution containing cell lysis buffer and trypsin is added to stop the deacetylation reaction and cleave the deacetylated substrate. The mixture is incubated at room temperature for 15 minutes. The fluorescence of released AMC is measured using a microplate reader with excitation at 355 nm and emission at 460 nm.
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| Cell Assay |
For cell-based HDAC activity assays, cells are seeded in 96-well tissue culture plates at a density of 6 × 10⁴ cells per 100 μL of medium and cultured overnight. The cells are then treated with the test compound or vehicle, followed by the addition of 25 μM Boc-Lys(Ac)-AMC. The plates are incubated at 37°C with 5% CO₂ for 2-3 hours. After incubation, the HDAC developer solution containing cell lysis buffer and trypsin is added, and the plates are incubated at room temperature for 15 minutes. Fluorescence is measured at Ex/Em = 355 nm/460 nm using a microplate reader.
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| Animal Protocol |
Specific in vivo animal experiment protocols for Boc-Lys(Ac)-AMC are not applicable as the compound is a substrate used in in vitro biochemical and cellular assays, not a therapeutic agent for animal studies. It is used as a research tool to measure HDAC activity in tissue samples or cell lysates derived from animal models. For such applications, tissues are collected from animals, lysed, and the lysates are incubated with Boc-Lys(Ac)-AMC to measure HDAC activity ex vivo.
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| ADME/Pharmacokinetics |
Specific pharmacokinetic data for Boc-Lys(Ac)-AMC are not applicable as the compound is a substrate used in biochemical assays rather than a therapeutic agent. It is not designed for systemic administration or pharmacological activity in vivo. The compound's properties as a small molecule (MW 445.51) and its cell permeability make it suitable for use in cell-based assays, but it is not intended for pharmacokinetic characterization as a drug candidate.
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| Toxicity/Toxicokinetics |
Specific toxicity data for Boc-Lys(Ac)-AMC are not applicable as the compound is a research tool used in biochemical assays, not a therapeutic agent. It is intended for laboratory use only and is not for human consumption or clinical applications. Standard laboratory safety precautions should be followed when handling this compound, including the use of appropriate personal protective equipment.
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| References | |
| Additional Infomation |
Boc-Lys(Ac)-AMC is a cell-permeable fluorometric HDAC substrate (Ex/Em = 355 nm/460 nm). Its chemical name is (S)-tert-butyl (6-acetamido-1-((4-methyl-2-oxo-2H-chromen-7-yl)amino)-1-oxohexan-2-yl)carbamate. The compound is designed for biological research and industrial applications and is not intended for individual clinical or medical purposes. It is typically stored at -20°C, sealed and away from moisture.
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| Molecular Formula |
C23H31N3O6
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|---|---|
| Molecular Weight |
445.51
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| Exact Mass |
445.221
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| CAS # |
233691-67-3
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| PubChem CID |
9846360
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| Appearance |
White to off-white solid powder
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| LogP |
4.094
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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 |
10
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| Heavy Atom Count |
32
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| Complexity |
743
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC1=CC(=O)OC2=C1C=CC(=C2)NC(=O)[C@H](CCCCNC(=O)C)NC(=O)OC(C)(C)C
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| InChi Key |
SUTRDULVNIPNLW-SFHVURJKSA-N
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| InChi Code |
InChI=1S/C23H31N3O6/c1-14-12-20(28)31-19-13-16(9-10-17(14)19)25-21(29)18(8-6-7-11-24-15(2)27)26-22(30)32-23(3,4)5/h9-10,12-13,18H,6-8,11H2,1-5H3,(H,24,27)(H,25,29)(H,26,30)/t18-/m0/s1
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| Chemical Name |
tert-butyl N-[(2S)-6-acetamido-1-[(4-methyl-2-oxochromen-7-yl)amino]-1-oxohexan-2-yl]carbamate
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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 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) |
DMSO : 100 mg/mL (224.46 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 | 2.2446 mL | 11.2231 mL | 22.4462 mL | |
| 5 mM | 0.4489 mL | 2.2446 mL | 4.4892 mL | |
| 10 mM | 0.2245 mL | 1.1223 mL | 2.2446 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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