Pt-Luc
Pt-Luc is a bioluminescent probe used for real-time, non-invasive monitoring of Pt (IV) prodrug reduction in living cells and animal models. The detection mechanism of Pt-Luc relies on the intracellular reduction mediated by active cysteine residues in proteins. Pt-Luc can penetrate the membrane of living cells; after entering cells, active cysteine thiolates in high-molecular-weight protein components reduce the Pt (IV) complex, thereby cleaving the caged aminoluciferin substrate. The released free aminoluciferin is then oxidized by luciferase in the presence of ATP to generate a bioluminescent signal, whose intensity is directly correlated with the reduction degree of Pt (IV).
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- Fòrmula: C20H21N5O9PtS2
- Peso molecular:734.62
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Almacenamiento:
Please store the product under the recommended conditions in the Certificate of Analysis.
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Actividad biológica
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Chemical Information
- Pureza y Documentación
- Referencias
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Help & FAQs
Actividad biológica
Descripciòn
In Vitro
Operating Instructions (The following is a recommended experimental protocol for guidance only; adjust according to specific requirements)
1. Stock Solution Preparation
1.1 Solvent: For most dyes, organic solvents are commonly used for dissolution, such as anhydrous DMSO.
1.2 Concentration Recommendation: It is generally recommended to prepare a high-concentration stock solution of 1-10 mM.
2. Working Solution Preparation
2.1 Diluent: Cell culture medium.
2.2 Working Concentrations: 0.5 μM, 5 μM, 6.25 μM, 10 μM, 12.5 μM, 20 μM, 25 μM, 50 μM.
2.3 Notes: Adjust the working solution concentration as needed; prepare fresh before use.
3. Staining Procedure
3.1 Sample Type: Adherent cancer cells expressing luciferase (A549-Luc2, MDA-MB-231-Luc)[1].
3.1.1 Adherent cells: Trypsin digestion is not required for imaging; cells need to be washed before incubation with the probe.
3.2 Incubation Conditions: Incubate with Pt-Luc working solution at 37°C for up to 180 min.
3.3 Washing Step: After pretreatment with Erastin (HY-15763) or
L-Buthionine-(S,R)-sulfoximine (HY-106376A), wash the cells once with buffer.
4. Control Group Setup
4.1 GSH Depletion Control Group: Pretreat cells with 250 μM L-Buthionine-(S,R)-sulfoximine for 24 h to deplete intracellular GSH.
4.2 Cysteine Starvation Control Group: Pretreat cells with 100 μM Erastin for 5 min followed by 24 h of incubation, or incubate cells in cystine-free medium.
4.3 Thiol Blocking Control Group: Pretreat cells with N-ethylmaleimide (HY-D0843) (3.125-50 μM) or Monobromobimane (HY-100041) (62.5-1000 μM) to block intracellular thiols.
4.4 Blank Control Group: Incubate cells with 0 μM Pt-Luc (cell culture medium only).
5. Detection and Analysis
5.1 Instrument: CCD camera for bioluminescence imaging.
5.2 Result Analysis:
5.2.1 Fluorescence Intensity Changes: Bioluminescence photon flux increases with rising Pt-Luc concentration and incubation time; signals decrease significantly in cysteine-starved or thiol-blocked cells, while no significant change occurs in GSH-depleted cells.
5.2.2 Fluorescence Localization: In in vivo models, bioluminescence localizes to tumor sites.
5.2.3 Color Change: Yellow bioluminescent signals are detectable after probe activation.
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
In Vivo
Pt-Luc (50 µM, 200 µL per 20 g mouse body weight; i.p.) shows bioluminescence reduced by 63% when thiols are blocked with NEM in murine lung cancer xenografts, confirming reactive cysteines in proteins are the dominant reductants for Pt(IV) prodrug activation in vivo[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:Balb/c nude mice[1]
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Dosage:50 µM, 200 µL per 20 g mouse body weight
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Administration:i.p.
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Result:Reached total photon flux of 2.533e+07 at 20 minutes post-injection.
Remained stable in blood during transport to systemic circulation and largely maintained integrity within 30 minutes.\nReduced bioluminescence signal from tumor sites by 63% in NEM-treated mice compared to untreated mice.
Achieved total photon flux integrated over 30 minutes of 1.7×108 in NEM-treated mice versus 4.6×108 in untreated mice.
Chemical Information
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Peso molecular 734.62
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Fòrmula C20H21N5O9PtS2
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SMILES
O=C(O[Pt]12(N[C@@H]3CCCC[C@H]3N1)(OC(C(O2)=O)=O)O)NC4=CC5=C(N=C(S5)C6=N[C@H](CS6)C(O)=O)C=C4
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Envío
Room temperature in continental US; may vary elsewhere.
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Almacenamiento
Please store the product under the recommended conditions in the Certificate of Analysis.
Pureza y Documentación
Referencias
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Pt-Luc Related Classifications
HY-D3436 Related Classifications
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