| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
PDKtide TFA is a substrate for PDK1. It binds to the active site of PDK1 and is phosphorylated by the kinase at a specific serine/threonine residue. This phosphorylation event can be monitored using radiolabeled ATP (gamma-32P-ATP) or by using specific antibodies that recognize the phosphorylated peptide. It can also be used to screen for PDK1 inhibitors.
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| ln Vitro |
In vitro, PDKtide TFA is used as a standard substrate in PDK1 kinase activity assays. It shows no direct antiproliferative activity itself but is used in biochemical and cell-based assays to measure PDK1 activity. Its phosphorylation status reflects PDK1 activity. PDKtide TFA can also be used to study the PDK1-mediated signaling pathway in various cell lines.
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| ln Vivo |
PDKtide TFA is not intended for in vivo use as a therapeutic agent, but PDK1 inhibitors screened using PDKtide TFA have been tested in animal models. For example, PDK1 inhibitors (e.g., PDK-IN-1 with IC50 0.03 uM for PDK1) have shown anti-tumor activity in mouse xenograft models. PDKtide TFA itself is used as a research tool for studying PDK1 biology and screening inhibitors.
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| Enzyme Assay |
For PDK1 kinase assays, PDK1 enzyme is incubated with PDKtide TFA (100-500 uM) in kinase buffer (e.g., 50 mM HEPES pH 7.5, 10 mM MgCl2, 1 mM DTT, 0.01% Tween 20) at 30degC for 30-60 minutes. The reaction is initiated by adding ATP (10-100 uM, including gamma-32P-ATP). The reaction is terminated by adding phosphoric acid to a final concentration of 1%. An aliquot is spotted onto P81 phosphocellulose paper, which is washed with 0.75% phosphoric acid and then with acetone. The incorporated radioactivity is measured by liquid scintillation counting. IC₅0 values for inhibitors are calculated.
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| Cell Assay |
For cell-based assays, PDK1-expressing cells (e.g., HEK293, HeLa, or cancer cell lines) are cultured in DMEM + 10% FBS at 37degC, 5% CO2. PDKtide TFA (1-100 uM) is not typically used directly in cell-based assays because it is a peptide that does not readily cross cell membranes. Instead, it can be delivered by microinjection or transfection. Alternatively, PDK1 activity is assessed by Western blotting using phospho-specific antibodies against downstream targets of PDK1 (e.g., phospho-AKT at T308).
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| Animal Protocol |
PDKtide TFA is not administered directly to animals. However, PDK1 inhibitors identified using PDKtide TFA can be tested in animal models. For example, tumor-bearing mice are administered PDK1 inhibitors (e.g., PDK-IN-1) orally or intraperitoneally at doses of 10-50 mg/kg daily for 14-28 days. Tumor volume is measured every 2-3 days. Blood samples are collected for PK analysis. Tissues are harvested for Western blotting to confirm PDK1 inhibition.
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| ADME/Pharmacokinetics |
PDKtide TFA is a peptide and is not administered in vivo. Its pharmacokinetics are not studied. For PDK1 inhibitors that are screened using PDKtide TFA, PK parameters vary depending on the inhibitor. For example, PDK-IN-1 has good oral bioavailability and a moderate half-life. PDKtide TFA is typically stored as a powder at -20degC for up to 3 years, or in solution at -80degC for up to 1 year.
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| Toxicity/Toxicokinetics |
PDKtide TFA itself has no toxicity because it is not administered to animals. However, the TFA counterion can cause mild irritation if inhaled or in contact with skin. For PDK1 inhibitors identified using PDKtide TFA, toxicity profiles vary. At therapeutic doses, PDK1 inhibitors have shown no significant toxicity in animal models. Higher doses may cause weight loss and hepatotoxicity.
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| References | |
| Additional Infomation |
PDKtide TFA is a biological active peptide used as a standard substrate for PDK1 kinase activity assays. It is used in enzyme kinetics, signal transduction research (PI3K/AKT pathway), and drug discovery (screening of PDK1 inhibitors). PDK1 is a master kinase in the PI3K/AKT signaling pathway, which is frequently dysregulated in cancer. PDK1 inhibitors have potential therapeutic applications in oncology.
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| Molecular Formula |
C212H314N54O66S3.XC2HF3O2
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|---|---|
| Molecular Weight |
4771.28 (free base)
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| Related CAS # |
PDKtide
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| Sequence |
Lys-Thr-Phe-Cys-Gly-Thr-Pro-Glu-Tyr-Leu-Ala-Pro-Glu-Val-Arg-Arg-Glu-Pro-Arg-Ile-Leu-Ser-Glu-Glu-Glu-Gln-Glu-Met-Phe-Arg-Asp-Phe-Asp-Tyr-Ile-Ala-Asp-Trp-CysKTFCGTPEYLAPEVRREPRILSEEEQEMFRDFDYIADWC
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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 |
| 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 (with sonication)
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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.) |
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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