| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
The primary molecular target of ATP-polyamine-biotin is the ATP-binding site of protein kinases. As an ATP analogue, the compound mimics ATP and binds to the kinase active site, where it acts as a cosubstrate. The compound's polyamine moiety enhances cell permeability, allowing it to penetrate cell membranes and access intracellular kinases. The biotin moiety enables the labeling and subsequent detection or purification of kinase substrates via streptavidin-based methods (e.g., Western blot, pull-down, mass spectrometry). By binding to kinases and being transferred to substrates, ATP-polyamine-biotin enables the identification and characterization of kinase-substrate interactions and the study of kinase signaling networks.
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| ln Vitro |
PKA kinase and myelin basic protein (MBP), the full-length protein substrate, were treated with ATP-polyamine-biotin (APB). Kinase was required for the detection of biotinylation. Additionally, in the presence of the kinase inhibitor staurosporine, in the absence of ATP-polyamine-biotin, or during acid incubation, MBP biotinylation is lost as a result of cleavage of the phosphoramidate bond. It is only in the presence of APB co-substrate that the biotinylated Kemp peptide product is detected [1].
In vitro studies demonstrate that ATP-polyamine-biotin is an efficient kinase cosubstrate with conversions and kinetics similar to those of other known ATP analogues. The compound is cell-permeable and can be used to label kinase substrates in live cells. In biochemical assays, ATP-polyamine-biotin is used as a substrate for purified kinases, enabling the biotinylation of kinase substrates. The biotinylated products can be detected by streptavidin-HRP in Western blot or captured on streptavidin beads for mass spectrometry-based identification. The compound's ability to label kinase substrates in a kinase-dependent manner makes it a powerful tool for studying kinase specificity, substrate identification, and inhibitor screening. ATP-polyamine-biotin shows a cytotoxicity profile that should be considered when used in cellular assays. In cellular studies, ATP-polyamine-biotin is used to promote biotin labeling of kinase substrates in live cells. The compound is cell-permeable and can be added to cell culture medium, where it enters cells and acts as a cosubstrate for endogenous kinases. Kinase substrates are biotinylated in a kinase-dependent manner, and the labeled proteins can be detected by Western blot using streptavidin-HRP or captured on streptavidin beads for identification by mass spectrometry. This approach enables the identification of kinase substrates, the study of kinase signaling pathways, and the screening of kinase inhibitors. The compound has been used in various cell types and has applications in phosphoprotein purification and analysis. ATP-polyamine-biotin is a valuable tool for chemical biology and proteomics research. |
| ln Vivo |
In vivo studies of ATP-polyamine-biotin are limited, as the compound is primarily used as a research tool for in vitro and cellular applications. However, the compound's cell-permeability suggests that it could potentially be used in animal models for studying kinase activity in vivo. In such studies, ATP-polyamine-biotin could be administered to animals, and tissues could be collected for analysis of biotinylated proteins. However, no extensive in vivo pharmacological or toxicological studies have been reported for this compound. The compound is primarily used for chemical biology and proteomics research applications.
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| Enzyme Assay |
For kinase assays, ATP-polyamine-biotin is used as a substrate for purified kinases in vitro. Purified kinase is incubated with ATP-polyamine-biotin (typically 0.1-100 µM) and a peptide or protein substrate in kinase assay buffer (e.g., 50 mM HEPES, pH 7.5, 10 mM MgCl₂, 1 mM DTT) at 30°C for 30-60 minutes. The reaction is terminated by addition of SDS-PAGE loading buffer or by heat inactivation. The reaction products are separated by SDS-PAGE and transferred to a membrane. Biotinylated products are detected by streptavidin-HRP and chemiluminescence. For kinase activity assays, the amount of biotinylation is quantified by densitometry. For kinetic studies, the Km for ATP-polyamine-biotin is determined by varying the concentration of the compound and measuring the rate of biotinylation. For inhibitor screening, kinases are incubated with ATP-polyamine-biotin in the presence of varying concentrations of test inhibitors, and the inhibition of biotinylation is measured.
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| Cell Assay |
For cellular labeling studies, cells (e.g., cancer cell lines, primary cells) are cultured in appropriate medium with 10% FBS and antibiotics. Cells are seeded in 6-well or 10-cm dishes and grown to 70-80% confluence. ATP-polyamine-biotin is dissolved in water or buffer and added to the culture medium at final concentrations (typically 10-100 µM). Cells are treated for 1-4 hours at 37°C. For kinase inhibitor studies, cells are pre-treated with kinase inhibitors for 30-60 minutes before addition of ATP-polyamine-biotin. After treatment, cells are washed with PBS and lysed in RIPA buffer or lysis buffer with protease inhibitors. Biotinylated proteins are detected by Western blot using streptavidin-HRP or captured on streptavidin-agarose beads for pull-down assays. For mass spectrometry identification, biotinylated proteins are enriched on streptavidin beads, eluted, and analyzed by LC-MS/MS. For phosphoprotein purification, biotinylated phosphoproteins are purified using streptavidin affinity chromatography.
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| Animal Protocol |
For in vivo studies, ATP-polyamine-biotin could be administered intravenously or intraperitoneally to mice at doses determined from preliminary toxicity studies. Tissues would be collected at various time points and processed for Western blot analysis or mass spectrometry to identify biotinylated proteins. However, such studies are not typical for this compound, and no specific protocols are reported in the literature.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for ATP-polyamine-biotin are limited, as the compound is used as a research reagent rather than a therapeutic agent. As a cell-permeable ATP analogue, the compound is expected to be taken up by cells and distributed to tissues. Its metabolism would involve hydrolysis of the phosphoanhydride bonds and degradation of the polyamine and biotin moieties. However, no dedicated pharmacokinetic studies have been reported. The compound is primarily used in vitro and in cellular applications and is not intended for in vivo pharmacokinetic characterization.
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| Toxicity/Toxicokinetics |
Toxicological data for ATP-polyamine-biotin are limited. The compound shows a cytotoxicity profile that should be considered when used in cellular assays. At the concentrations used for labeling (typically 10-100 µM), the compound may have some cytotoxic effects depending on the cell type and exposure time. No acute toxicity, organ-specific toxicity, or mutagenicity data have been reported. As with all chemical reagents, appropriate safety precautions should be taken when handling ATP-polyamine-biotin, including the use of personal protective equipment (gloves, lab coat, safety glasses) and work in a well-ventilated area.
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| References | |
| Additional Infomation |
ATP-polyamine-biotin (CAS 1800401-93-7) is the first cell-permeable ATP analogue and an efficient kinase cosubstrate. It binds to ATP-binding sites of protein kinases and promotes biotin labeling of kinase substrates in live cells. The compound has conversions and kinetics similar to other ATP analogues. Its molecular formula is C₃₂H₅₈N₁₁O₁₄P₃S with a molecular weight of 945.85. ATP-polyamine-biotin is a powerful tool for studying kinase signaling, identifying kinase substrates, and discovering kinase inhibitors. It has applications in phosphoprotein purification and analysis. The compound is strictly for research use only.
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| Molecular Formula |
C32H58N11O14P3S
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|---|---|
| Molecular Weight |
945.8545
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| Exact Mass |
945.309
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| CAS # |
1800401-93-7
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| PubChem CID |
118704767
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| Appearance |
White to off-white solid powder
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| Density |
1.8±0.1 g/cm3
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| Index of Refraction |
1.730
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| LogP |
-3.53
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| Hydrogen Bond Donor Count |
10
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| Hydrogen Bond Acceptor Count |
22
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| Rotatable Bond Count |
27
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| Heavy Atom Count |
61
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| Complexity |
1590
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| Defined Atom Stereocenter Count |
7
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| SMILES |
CN(CCCCN(C)CCCNP(=O)(O)OP(=O)(O)OP(=O)(O)OC[C@@H]1[C@H]([C@H]([C@@H](O1)N2C=NC3=C(N=CN=C32)N)O)O)CCCNC(=O)CCCC[C@H]4[C@@H]5[C@H](CS4)NC(=O)N5
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| InChi Key |
MODMQMYTNMWWCS-MCINONILSA-N
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| InChi Code |
InChI=1S/C32H58N11O14P3S/c1-41(15-7-11-34-24(44)10-4-3-9-23-25-21(18-61-23)39-32(47)40-25)13-5-6-14-42(2)16-8-12-38-58(48,49)56-60(52,53)57-59(50,51)54-17-22-27(45)28(46)31(55-22)43-20-37-26-29(33)35-19-36-30(26)43/h19-23,25,27-28,31,45-46H,3-18H2,1-2H3,(H,34,44)(H,50,51)(H,52,53)(H2,33,35,36)(H2,38,48,49)(H2,39,40,47)/t21-,22+,23-,25-,27+,28+,31+/m0/s1
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| Chemical Name |
N-[3-[4-[3-[5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]propyl-methylamino]butyl-methylamino]propyl]-[[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-hydroxyphosphoryl]oxyphosphonamidic acid
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 : ~6 mg/mL (~6.34 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 7.69 mg/mL (8.13 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with heating and sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.0573 mL | 5.2863 mL | 10.5725 mL | |
| 5 mM | 0.2115 mL | 1.0573 mL | 2.1145 mL | |
| 10 mM | 0.1057 mL | 0.5286 mL | 1.0573 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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