Purmorphamine [483367-10-8]
Cat# T1810-5mg
Size : 5mg
Brand : TargetMol
Purmorphamine
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Purity:99.48%
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COA HPLC HNMR
Purmorphamine
Catalog No. T1810Cas No. 483367-10-8
Purmorphamine (Shh Signaling Antagonist VI), which directly binds and activates Smoothened, blocks BODIPY-cyclopamine binding to Smo. It also is an inducer of osteoblast differentiation.
All TargetMol products are for research purposes only and cannot be used for human consumption. We do not provide products or services to individuals. Please comply with the intended use and do not use TargetMol products for any other purpose. Pack Size | ||
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5 mg | In Stock | |
10 mg | In Stock | |
50 mg | In Stock | |
1 mL x 10 mM (in DMSO) | In Stock |
Product Introduction
Bioactivity
Chemical Properties
Storage & Solubility Information
Description | Purmorphamine (Shh Signaling Antagonist VI), which directly binds and activates Smoothened, blocks BODIPY-cyclopamine binding to Smo. It also is an inducer of osteoblast differentiation. |
Targets&IC50 | Smo:1.5 μM |
In vitro | In rat models, based on human mesenchymal stem cells, Purmorphamine upregulates the expression of ALP. |
In vivo | In multipotent C3H10T1/2 cells, Purmorphamine can induce osteogenesis with an EC50 of 1 μM. It directly binds to Smoothened, thereby activating the Hedgehog pathway with an IC50 of 1.5 μM. |
Kinase Assay | Binding assay: Smo binding assays are conducted with BODIPY-cyclopamine and Smo-overexpressing cells as previously described4,5, using CMV promoter-based, SV40 origin-containing expression constructs for Smo-Myc3, the deletion mutant Smo?CRD (deletion of amino acids 68 to 182), and Smo?CT (deletion of amino acids 556 to 793). HEK 293T cells are grown on poly-D-lysine-treated glass coverslips in 12-well plates until 70% confluency and then transfected with the appropriate expression construct (0.5 ?g/well) using FuGene 6 according the manufacturer's protocols. Two days after transfection, the HEK 293T cells are incubated with DMEM containing 0.5% bovine calf serum, 5 nM BODIPY-cyclopamine, and varying concentrations of Purmorphamine (0, 1.5, or 5 ?M) (1 mL/well) for 1 h at 37 ℃. The Smo-overexpressing cells are then washed with 1 × PBS buffer (1 mL/well), mounted with DAPI-containing medium, and visualized using a Leica DM4500B fluorescence microscope. For binding assays using fixed cells, the Smo-overexpressing HEK 293T cells are fixed with 3% paraformaldehyde in 1 × PBS buffer for 10 min at room temperature (1 mL/well), treated with 1 × PBS containing 10 mM glycine and 0.2% sodium azide for 5 min (1 mL/well), washed with 1 × PBS buffer (1 mL/well), and treated with the Purmorphamine-containing media described above for 4 h at room temperature. |
Cell Research | C3H10T1/2 cells are expanded in T175 flasks; cells at 13th passage are detached by trypsin/EDTA and diluted in the growth media. The resulting cell suspension is then plated into black clear bottom 384-well plates with 2500 cells/well in 100 μL growth medium using a Multi-dropTM liquid delivery system. After overnight incubation, cells attached to the bottom of the wells. A stock solution of each Purmorphamine in DMSO (500 nL) is delivered into corresponding well using a Mini TrakTM multiposition dispenser system to make a final concentration of 5 μM of Purmorphamine. Cells are then incubated at 37 ℃ with 5% CO2 in air atmosphere. After 4 days, the medium is removed and 10 μL of passive lysis buffer is added into each well. After 5 min, 10 μL of alkaline phosphatase substrate solution is added to each well. After incubating 15 min at room temperature, the plates are read on an Acquest high-throughput plate reader following the manufacturer's protocol.(Only for Reference) |
Alias | Shh Signaling Antagonist VI |
Molecular Weight | 520.62 |
Formula | C31H32N6O2 |
Cas No. | 483367-10-8 |
Storage | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. | |||||||||||||||||||||||||||||||||||
Solubility Information | DMSO: 55 mg/mL (105.64 mM) | |||||||||||||||||||||||||||||||||||
Solution Preparation Table | ||||||||||||||||||||||||||||||||||||
DMSO
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In Vivo Formulation Calculator (Clear solution)
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Mother liquor preparation method: 2 mg of drug dissolved in 50 μL DMSO
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Preparation method for in vivo formula: Take 50 μL DMSO
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Keywords
inhibitAutophagySmoSmoothenedInhibitorPurmorphamine
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