| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 100mg | |||
| Other Sizes |
| Targets |
Histone deacetylases (HDACs), particularly class I and class II HDACs, as well as sirtuins (class III HDACs) and lysine-specific proteases. Ac-Lys-AMC serves as a fluorescent substrate that mimics acetylated lysine residues in histone proteins. When HDACs remove the acetyl group from the lysine side chain, the deacetylated product becomes susceptible to cleavage by trypsin or other proteases, which releases the highly fluorescent AMC group. This two-step enzymatic reaction enables sensitive and real-time monitoring of HDAC and deacetylase activities in biochemical assays.
|
|---|---|
| ln Vitro |
In vitro, Ac-Lys-AMC functions as a fluorogenic substrate for measuring histone deacetylase (HDAC) activity. It is used in enzymatic assays to determine the activity of various HDAC isoforms including HDAC1, HDAC2, HDAC3, SIRT1, SIRT3, SIRT6, and SIRT7. Typical assay conditions use 250 uM Ac-Lys(Ac)-AMC substrate with 0.225 ug/well HDAC1-2 or 0.075 ug/well HDAC3. The compound enables sensitive detection of enzymatic kinetics with fluorescence readout upon AMC release. It is also used to evaluate the inhibitory potency of HDAC inhibitors in drug discovery platforms.
|
| ln Vivo |
In vivo studies of Ac-Lys-AMC are not typically performed as the compound is used exclusively as an in vitro biochemical research tool rather than a therapeutic agent. The substrate is designed for cell-free enzymatic assays and is not administered to animals or humans for pharmacological evaluation. Its applications are limited to measuring HDAC and deacetylase activities in tissue lysates, cell extracts, or purified enzyme preparations ex vivo.
|
| Enzyme Assay |
In vitro enzyme assays using Ac-Lys-AMC typically employ a two-step protocol. First, the substrate is incubated with the target enzyme (e.g., HDAC or deacetylase) in assay buffer at 37degC for 30-60 minutes to allow deacetylation. Second, a developer solution containing trypsin or Lys-C is added to cleave the deacetylated substrate and release the fluorescent AMC moiety. Fluorescence is measured at excitation ~360 nm and emission ~460 nm using a microplate reader. IC50 values for inhibitors are calculated from dose-response curves. The assay is compatible with high-throughput screening formats.
|
| Cell Assay |
In vitro cell-based assays using Ac-Lys-AMC are performed with cell lysates or nuclear extracts containing HDAC enzymes. Cells are lysed in HDAC assay buffer containing 50 mM NaCl, 50 mM Tris-HCl pH 7.5, 5% glycerol, and 0.3% Triton X-100 at 4degC for 20 minutes. Cleared lysates are incubated with Ac-Lys-AMC substrate (typically 200-250 uM) at 37degC for 30-60 minutes. Following deacetylation, developer solution is added and fluorescence is measured. The assay can be used to measure endogenous HDAC activity in response to drug treatments or genetic manipulations.
|
| Animal Protocol |
In vivo animal studies are not applicable for Ac-Lys-AMC as it is a research-use fluorogenic substrate rather than a pharmacological agent. The compound is not intended for therapeutic use or in vivo administration in animal models. Its applications are strictly limited to in vitro biochemical and cell-based assays for studying HDAC and deacetylase enzyme activities.
|
| ADME/Pharmacokinetics |
Pharmacokinetic properties of Ac-Lys-AMC are not applicable as the compound is used exclusively as an in vitro research tool. With a molecular weight of 345.39 and a molecular formula of C18H23N3O4, the compound is soluble in DMSO. It is typically stored as a powder at -20degC for up to 3 years or in solution at -80degC for up to 1 year. For short-term storage, 2-8degC is also suitable. As a substrate, pharmacokinetic parameters such as absorption, distribution, metabolism, and excretion are not determined.
|
| Toxicity/Toxicokinetics |
Ac-Lys-AMC is intended for research use only and is not for human therapeutic or diagnostic use. Standard safety precautions for handling chemical compounds apply. The compound is not approved for clinical use. No significant toxicity has been reported for this research substrate.
|
| Additional Infomation |
Ac-Lys-AMC is a research-grade fluorogenic substrate for histone deacetylases (HDACs) and deacetylases. Its synonyms include Hexanamide, (S)-2-Acetamido-6-amino-N-(4-methyl-2-oxo-2H-chromen-7-yl)hexanamide, and N2-Acetyl-N-(4-methyl-2-oxo-2H-chromen-7-yl)-L-lysinamide. Purity is typically ≥98%. The compound is widely used in epigenetic research, post-translational modification analysis, and drug discovery platforms. It enables sensitive detection of enzymatic activities and facilitates high-throughput screening of HDAC inhibitors. Not approved for clinical use.
|
| Molecular Formula |
C18H23N3O4
|
|---|---|
| Molecular Weight |
345.39292
|
| Exact Mass |
345.169
|
| CAS # |
156661-42-6
|
| PubChem CID |
15816975
|
| Appearance |
White to off-white solid powder
|
| LogP |
3.863
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
7
|
| Heavy Atom Count |
25
|
| Complexity |
546
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
CC1=CC(OC2=C1C=CC(NC([C@@H](NC(C)=O)CCCCN)=O)=C2)=O
|
| InChi Key |
NBOQAQRQQNJREL-HNNXBMFYSA-N
|
| InChi Code |
InChI=1S/C18H23N3O4/c1-11-9-17(23)25-16-10-13(6-7-14(11)16)21-18(24)15(20-12(2)22)5-3-4-8-19/h6-7,9-10,15H,3-5,8,19H2,1-2H3,(H,20,22)(H,21,24)/t15-/m0/s1
|
| Chemical Name |
(2S)-2-acetamido-6-amino-N-(4-methyl-2-oxochromen-7-yl)hexanamide
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~289.53 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.24 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 (7.24 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (7.24 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.8953 mL | 14.4764 mL | 28.9528 mL | |
| 5 mM | 0.5791 mL | 2.8953 mL | 5.7906 mL | |
| 10 mM | 0.2895 mL | 1.4476 mL | 2.8953 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
