| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Sakamototide substrate peptide TFA targets the catalytic domain of AMPK family kinases, serving as a specific peptide substrate. It is not an inhibitor or drug target but rather a molecular tool that mimics the phosphorylation site of natural AMPK substrates. The peptide is recognized by the active site of the kinase, allowing researchers to measure its enzymatic activity in vitro by monitoring the transfer of a phosphate group from ATP to the peptide.
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| ln Vitro |
Sakamototide substrate peptide TFA is used in in vitro kinase activity assays to measure the function of AMPK family members. It can be employed to determine kinetic parameters such as the Michaelis constant (Km) and maximum velocity (Vmax). It is also used in high-throughput screening (HTS) formats to identify and evaluate small-molecule modulators (inhibitors or activators) of AMPK activity, which is a key target for metabolic disorders like diabetes and obesity.
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| ln Vivo |
The peptide is not intended for in vivo use; it is an in vitro biochemical tool. Its purpose is to quantify AMPK activity in cell lysates or with purified enzymes. As a substrate, it does not possess intrinsic in vivo activity. It is used to study the regulation of AMPK in cellular signaling pathways, and the data generated from these in vitro assays can inform in vivo models of metabolism.
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| Enzyme Assay |
A cell-free AMPK kinase assay is performed using Sakamototide substrate peptide TFA. The reaction mixture (25-50 uL) contains kinase buffer (e.g., 50 mM HEPES, pH 7.5, 5 mM MgCl2, 1 mM DTT), 50-100 microM ATP (including gamma-32P-ATP for radioactive assays or unlabeled ATP for luminescent assays), and 100-500 microM of the peptide substrate. Purified AMPK (0.1-1 microg) is added to start the reaction. After incubation at 30degC for 30-60 min, the reaction is terminated by adding 0.5% phosphoric acid. The reaction mixture is spotted onto P81 phosphocellulose paper, which binds the phosphorylated peptide. The papers are washed, and the incorporated radioactivity is quantified by scintillation counting. For non-radioactive detection, an ADP-Glo™ assay can be used.
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| Cell Assay |
Sakamototide substrate peptide TFA can be used to measure AMPK activity in cellular lysates. Cells (e.g., HEK293 or HepG2 cells) are grown to 80-90% confluency and treated with AMPK activators (e.g., metformin, AICAR) or inhibitors. After treatment, cells are lysed in a non-denaturing lysis buffer with protease and phosphatase inhibitors. The cleared lysate (10-50 ug protein) is incubated with the peptide substrate in a reaction mixture containing ATP for 30-60 min at 30degC. The AMPK activity is measured by quantifying the phosphorylation of the substrate using a phospho-specific antibody in an ELISA format or by LC-MS/MS.
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| Animal Protocol |
Sakamototide substrate peptide TFA is not administered in vivo. However, to assess AMPK activity ex vivo in tissues, a standard protocol involves animal treatment (e.g., mice injected with metformin) followed by tissue collection (e.g., liver, skeletal muscle). Tissue samples are homogenized in lysis buffer, and the cleared homogenate is used in the kinase activity assay as described in the cell-based protocol section. The peptide substrate allows for the sensitive measurement of endogenous AMPK activity in these tissue lysates.
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| ADME/Pharmacokinetics |
Sakamototide substrate peptide TFA has a molecular formula of C₇1H123F3N30O2₅ and a molecular weight of 1853.92. The peptide should be stored as a lyophilized powder at -20degC for up to 3 years, sealed and away from moisture. In solution, it is stable for 6 months at -80degC or 1 month at -20degC. The TFA salt ensures solubility in aqueous buffers for biochemical assays.
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| Toxicity/Toxicokinetics |
No specific toxicity data are available for the peptide substrate. As a research-grade peptide, it is not intended for in vivo use and is expected to have low inherent toxicity. Standard laboratory safety precautions for handling peptides should be followed, including the use of gloves and eye protection to avoid accidental exposure. The TFA counterion is considered safe at the low levels used in these assays.
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| References |
[1]. Banerjee S,et al. Characterization of WZ4003 and HTH-01-015 as selective inhibitors of the LKB1-tumour-suppressor-activated NUAK kinases. Biochem J. 2014 Jan 1;457(1):215-25.
[2]. Banerjee S, et al. Interplay between Polo kinase, LKB1-activated NUAK1 kinase, PP1βMYPT1 phosphatasecomplex and the SCFβTrCP E3 ubiquitin ligase. Biochem J. 2014 Jul 15;461(2):233-45. |
| Additional Infomation |
Sakamototide substrate peptide TFA is a research-grade peptide and is not approved for clinical use. It is a synthetic peptide substrate for the AMPK family of kinases, widely used in basic research for signal transduction studies and drug discovery. The TFA salt form is the standard for research applications, enhancing solubility and stability for in vitro biochemical assays. This product is for research use only and not for human diagnostic or therapeutic applications.
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| Molecular Formula |
C71H123F3N30O25
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| Appearance |
White to off-white solid powder
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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, 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) |
H2O :~100 mg/mL (~53.94 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 50 mg/mL (26.97 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
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
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