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Daphnetin (7,8-dihydroxycoumarin), one coumarin derivative isolated from plants of the Genus Daphne, is a protein kinase inhibitor, with IC50s of 7.67 μM, 9.33 μM and 25.01 μM for EGFR, PKA and PKC in vitro, respectively. It is also a chelator and an antioxidant. Daphnetin causes a 50% inhibition (IC50) of 3H-hypoxanthine incorporation by Plasmodium falciparum at concentrations between 25 and 40 uM.
| Cell Experiment | |
|---|---|
| Cell lines | MCF-7 cells |
| Preparation method | Using the microculture MTT assay to estimate the cytostatic effect of Daphnetin which tested on the MCF-7 tumor cells . The assay is based on the reduction of soluble tetrazolium salt by mitochondria of viable cells. The reduced product,which is an insoluble purple-colored formazan, is dissolved in dimethyl sulfoxide and measured spectrophotometrically (570 nm). The amount of formazan formed is proportional to the number of viable cells. Cells (3 × 103) are seeded in each of the 96 microplate wells in a 200 μL medium which contains the corresponding concentration of daphnetin. Testing daphnetin at five concentrations (12.5 μM, 25 μM, 50 μM, 100 μM, 200 μM). 24-hour, 48-hour, and 72-hour exposure later , the percentage of proliferative inhibition of treated cells is estimated against the solvent-treated control cells (PI% = [(T/C) − 1] × 100). PI = proliferation inhibition; T = treated, C = control. Calculating the IC50 from the least square concentration-response regressions |
| Concentrations | 12.5 μM, 25 μM, 50 μM, 100 μM, 200 μM |
| Incubation time | 24 hours, 48 hours, and 72 hours |
| Animal Experiment | |
|---|---|
| Animal models | Immature CD1 (Im) female mice |
| Formulation | Corn oil |
| Dosages | 35 mg/kg, 70 mg/kg, and 140 mg/kg |
| Administration | Subcutaneously |
| Molecular Weight | 178.14 |
| Formula | C9H6O4 |
| CAS Number | 486-35-1 |
| Solubility (25°C) | DMSO 30 mg/mL |
| Storage |
Powder -20°C 3 years ; 4°C 2 years In solvent -80°C 6 months ; -20°C 1 month |
| Related PKC Products |
|---|
| Sapintoxin D
Sapintoxin D is a fluorescent phorbol ester that can be used to study the activation of protein kinase C (PKC). |
| Bryostatin 3
Bryostatin 3, a macrocyclic lactone, is a protein kinase C activator, with a Ki of 2.75 nM. |
| Ruboxistaurin mesylate
Ruboxistaurin (LY333531) mesylate is an orally active, selective and ATP competitive PKCβ inhibitor with IC50 values of 4.7 and 5.9 nM for PKCβI and PKCβII, respectively. |
| PKC β pseudosubstrate
PKC β pseudosubstrate is a selective cell-permeable inhibitor of PKC. |
| Protein Kinase C (19-31)
Protein Kinase C (19-31), a peptide inhibitor of protein kinase C (PKC), is used as protein kinase C substrate peptide for testing the protein kinase C activity. |
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