PTPA-QM
PTPA-QM is a low-cytotoxicity Aβ protein probe that can be used for live-cell imaging and tissue section staining (Lys. Ex/Em = 385/615 nm). PTPA-QM can intercalate into the β-sheet layered structure of β-amyloid fibrils, form intermolecular interactions with amino acid residues, restrict intramolecular rotation and trigger fluorescence activation for imaging purposes. PTPA-QM is applicable to Alzheimer's disease-related research. Maximum excitation/emission wavelength: 448/605 nm.
Nur für Forschungszwecke. Wir verkaufen nicht an Patienten.
- Formel: C38H27N5
- Molecular Weight:553.65
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Speicherung:
Please store the product under the recommended conditions in the Certificate of Analysis.
Biologische Aktivität
Beschreibung
In Vitro
Guidelines (The following is our recommended experimental protocol. This protocol is for reference only; specific operations should be adjusted according to your actual needs.)
1. Preparation of PTPA-QM Working Solution
1.1 Preparation of Stock Solution
Prepare 1 mM PTPA-QM stock solution using DMSO. Aliquot and store at -20°C protected from light, avoiding repeated freeze-thaw cycles.
1.2 Preparation of Working Solution
Dilute the stock solution with PBS buffer to prepare 10 μM PTPA-QM working solution.
Note: Please adjust the working solution concentration according to your actual situation, and prepare fresh solution each time, protecting from light throughout the process.
2. Cell Staining
2.1 Cell Preparation
2.2 Add PTPA-QM working solution and incubate for 60 min.
2.3 Wash cells with PBS and detect them under a fluorescence microscope or flow cytometer.
3. Staining of sections
3.1 Wash brain tissue sections three times with PBS.
3.2 Add PTPA-QM working solution (2 μM) and incubate at room temperature in the dark for 3 min.
3.3 Wash stained brain tissue sections three times with DMSO:H2O = 1:1.
PTPA-QM (10 μM) is a highly efficient, selective, and photostable fluorescence probe for viscosity detection. Its fluorescence intensity in 99% glycerol is 22 times higher than that in pure DMSO, and it is minimally affected by biologically related substances. [1]
PTPA-QM (10 μM) exhibits viscosity-dependent fluorescence enhancement, with its fluorescence intensity increasing approximately 8-fold in high-viscosity glycerol compared to low-viscosity PBS; simultaneously, this probe possesses aggregation-induced emission properties[3].
PTPA-QM (0.30-40.00 Mm, 24 h) shows low cytotoxicity to PC12 cells[1].
PTPA-QM (10 μM, 1 h) can specifically bind to Aβ fibrils through various intermolecular interactions, and its fluorescence intensity increases approximately 2-fold after binding in cell-free experiments[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:PC12 cells
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Concentration:10 μM
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Incubation Time:1 h
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Result:Produced strong red fluorescence visible via CLSM after incubation for 1 h, indicating successful cell penetration and intracellular fluorescence imaging.
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Cell Line:PC12 cells
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Concentration:0.3 μM, 0.6 μM, 1.25 μM, 2.5 μM, 5 μM, 10 μM, 20 μM, 40 μM
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Incubation Time:24 h
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Result:Exhibited low cytotoxicity to PC12 cells.
Chemical Information
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Molecular Weight 553.65
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Formel C38H27N5
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SMILES
CN(C1=C/2C=CC=C1)C(/C=C/C(C=C3)=CC=C3N(C4=CC=CC=C4)C(C=C5)=CC=C5C6=CC=NC=C6)=CC2=C(C#N)/C#N
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Versand
Room temperature in continental US; may vary elsewhere.
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Speicherung
Please store the product under the recommended conditions in the Certificate of Analysis.
Protokoll
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Cell Cytotoxicity Assay
Cytotoxicity assays are usually based on the assessment of cell membrane damage, which can also be indirectly detected by measuring cell viability. Detection methods include MTT assay, CKK-8 assay, LDH assay and ATP assay, etc.
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Mammalian live/dead viability and cytotoxicity staining
Live/dead viability and cytotoxicity staining assays are based on the simultaneous detection of intracellular esterase activity in metabolically active (viable) cells and membrane integrity loss in non-viable cells. In commonly used dual-staining approaches, membrane-permeant fluorogenic substrates are converted by intracellular esterases into fluorescent products in live cells, while impermeant DNA-binding dyes selectively enter cells with compromised plasma membranes and label nucleic acids in dead or dying cells, enabling discrimination between viable and non-viable populations by fluorescence microscopy or flow cytometry.
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Paraffin Section
The tissue is embedded in paraffin wax, cut into thin slices by a microtome, stained and sealed and finally made into samples for microscopic observation.
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Amyloid: Congo Red Amyloid Staining
Congo red amyloid staining is a histochemical method used to detect extracellular amyloid deposits in tissue sections based on the affinity of Congo red dye for β-pleated sheet-rich protein aggregates. When bound to amyloid, Congo red produces characteristic apple-green birefringence under polarized light microscopy, which is widely regarded as a diagnostic feature of amyloid deposition in histopathology. The diagnostic principle relies on the combination of dye binding (congophilia) and optical anisotropy under polarized illumination, which distinguishes amyloid from most non-amyloid eosinophilic extracellular deposits in routine histological evaluation. Amyloid identification by Congo red staining remains a cornerstone in diagnostic pathology despite the availability of adjunct methods such as immunohistochemistry and mass spectrometry, particularly because of its ability to localize deposits directly within tissue architecture. The specificity of Congo red-positive deposits is incre
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Cell Viability Determination by MTT Colorimetric Assay
The following protocol uses the MTT colorimetric assay as a classic literature-established method for assessing cell viability/metabolic activity in cultured mammalian cells. MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] is reduced by metabolically active cells to a colored formazan product; the amount of formazan is quantified spectrophotometrically and provides an indirect measure of metabolically active viable cells. Importantly, MTT reduction reflects cellular oxidoreductase/metabolic activity rather than an absolute direct count of living cells, so changes in cellular metabolism can alter the signal independently of cell number.
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Alzheimer’s Disease Modeling
Alzheimer’s Disease (AD) is a neurodegenerative disorder characterized by a progressive decline in cognitive functions and loss of specific types of neurons and synapses. Alzheimer's symptoms can be simulated in mice by injecting drugs (such as Aβ) or genetically modified.
Reinheit & Dokumentation
Verweise
[1]. Fang Y, et al. A Novel Aggregation-Induced Emission Fluorescent Probe for Detection of β-Amyloid Based on Pyridinyltriphenylamine and Quinoline-Malononitrile. Biosensors (Basel). 2023;13(6):610. Published 2023 Jun 2. [Content Brief]
[2]. Zheng M, et al. Fluorescent Materials with Excellent Biocompatibility and Their Application in Bio-Sensing, Bio-Imaging. Biosensors (Basel). 2023;13(10):906. Published 2023 Sep 26. [Content Brief]
Calculators
Konzentration (Stammlösung) × Volumen (Stammlösung) = Konzentration (Ziellösung) × Volumen (Ziellösung)