Spring red
Spring Red is a fluorescent probe for mitochondrial pH detection (Ex/Em = 530/680 nm) that can be taken up by live cells and mitochondria. The detection mechanism of Spring Red depends on pH changes and mitochondrial membrane potential. Fluorescence intensity increases with elevated pH, and a significant pH-dependent fluorescent response is observed in the pH range of 5.0‑8.0. Spring Red is suitable for long-term live-cell imaging and in vivo imaging, and can be used to monitor dynamic mitochondrial pH changes.
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- No. CAS: 1616505-01-1
- Fòrmula: C31H35ClN2O
- Peso molecular:487.08
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Almacenamiento:
Please store the product under the recommended conditions in the Certificate of Analysis.
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Actividad biológica
Operating Instructions
(The following is a recommended experimental protocol for guidance only and should be adjusted according to your specific requirements).
1. Stock Solution Preparation
1.1 Solvent: Universal buffer (0.1 M citric acid, 0.1 M KH2PO4, 0.1 M Na2B4O7, 0.1 M Tris, 0.1 M KCl).
1.2 Concentration recommendation: 1 mM.
2. Working Solution Preparation
2.1 Working concentration: 30 μM.
2.2 Notes: Adjust the working solution concentration as needed; prepare and use immediately.
3. Staining Procedure
3.1 Sample type[1]:
3.1.1 Adherent cells: Human hepatocellular carcinoma cells (HepG2).
3.2 Incubation conditions: Incubate adherent cells with 30 μM Spring Red for 10 min.
3.3 Washing step: Wash away lactic acid and pyruvate from HepG2 cells to reverse mitochondrial pH changes.
4. Control Setup
4.1 Localization imaging control: Stain HepG2 cells with Mito Green as a reference dye for mitochondrial localization.
4.2 Biological interferent control group: Test the interference of various common cations and ROS on the fluorescence signal of Spring Red in the universal buffer system to verify the anti-interference performance of the probe.
5. Detection and Analysis
5.1 Instrument: A laser confocal fluorescence microscope is used for cell imaging.
5.1.1 Excitation/emission wavelength: λex = 543 nm, λem = 650-800 nm.
5.2 Result analysis:
5.2.1 Fluorescence intensity change: Fluorescence intensity increases with elevated pH and decreases with reduced pH.
5.2.2 Fluorescence localization: Accumulates in the mitochondria of HepG2 cells.
5.2.3 Color change: Emits red near-infrared fluorescence.
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Cell Line:HL‑7702 cells
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Concentration:1.0×10‑4 M, 1.0×10‑5 M, 1.0×10‑6 M, 1.0×10‑7 M , 1.0×10‑8 M
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Incubation Time:12 h
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Result:Calculated IC50 value of Spring Red was 4.8×10‑3 M, Spring Red exhibited low cytotoxicity under cell imaging experimental conditions.
(The following is a recommended experimental protocol for guidance only, and adjustments are required based on your specific needs.)
1. Stock Solution Preparation
1.1 Solvent: Universal buffer (0.1 M citric acid, 0.1 M KH2PO4, 0.1 M Na2B4O7, 0.1 M Tris, 0.1 M KCl).
1.2 Recommended concentration: 1 mM.
2. Working Solution Preparation
2.1 Working concentration: 50 μM (for mice).
2.2 Notes: Adjust the working solution concentration as needed; prepare and use immediately.
3. Staining Procedure
3.1 Sample types[1]:
3.1.1 In vivo samples: Zebrafish, mice.
3.2 Incubation conditions: Inject 50 μM Spring Red intraperitoneally into mice.
4. Control Setup
4.1 In vivo imaging controls for mice:
4.1.1 LPS model control mice: Mice injected intraperitoneally with LPS only, without Spring Red probe treatment.
4.1.2 Probe control mice: Mice injected intraperitoneally with Spring Red probe only, without establishment of the intraperitoneal LPS-induced inflammation model.
4.2 In vivo imaging controls for zebrafish:
4.2.1 Positive organelle labeling control: Zebrafish stained with Mito Green, which serves as the positive reference dye for mitochondrial localization.
5. Detection and Analysis
5.1 Instruments: Laser confocal fluorescence microscope is used for zebrafish imaging; in vivo fluorescence imaging system is used for mouse imaging.
5.1.1 Excitation/emission wavelengths: For zebrafish imaging: λex = 543 nm, λem = 650-800 nm; for mouse imaging: λex = 530 nm, λem = 670 nm.
5.2 Result analysis:
5.2.1 Fluorescence intensity change: Fluorescence intensity increases with elevated pH and decreases with reduced pH.
5.2.2 Fluorescence localization: Enriches in the heart region of zebrafish; localizes in the abdominal cavity of mice.
5.2.3 Color change: Emits red near-infrared fluorescence.
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Chemical Information
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No. CAS 1616505-01-1
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Peso molecular 487.08
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Fòrmula C31H35ClN2O
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SMILES
CCN1C2=CC=CC=C2C(C)(/C1=C\C=C3C(Cl)=C(CCC\3)/C=C/C(C4=CC=C(C=C4)N(C)C)=O)C
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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)
Spring red Related Classifications
HY-D3132 Related Classifications
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