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LY333531 (Ruboxistaurin) hydrochloride competitively and reversibly inhibits PKCβI and PKCβII (IC50 values are 4.7 and 5.9 nM respectively). Selective for PKCβ over other PKC isozymes (IC50 values are 0.052, 0.25, 0.30, 0.36, 0.60 and >100 μM for PKCη, -δ, -γ, -α, -ε and -ζ respectively). LY333531 exhibits ATP dependent competitive inhibition of PKC beta I and is selective for PKC in comparison to other ATP dependent kinases (protein kinase A, calcium calmodulin, caesin kinase, src tyrosine kinase).
J Mol Cell Cardiol. 2015 Oct;138-47.
Irisin improves endothelial function in type 2 diabetes through reducing oxidative/nitrative stresses.
LY333531 hydrochloride purchased from AbMole
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Source | Journal of Molecular and Cellular Cardiology (2015). Figure 7.LY333531 (Abmole Company, Shanghai, China) |
| Method | Western blot | |
| Cell Lines | HUVECs | |
| Concentrations | 5 µM | |
| Incubation Time | 24 h | |
| Results | We found that PKC-β inhibitor LY333531 significantly suppressed augmented gp91phox expression. |
| Cell Experiment | |
|---|---|
| Cell lines | HUVECs |
| Preparation method | Western blot Aortas and HUVECs were lysed in lysis buffer or membrane and cytosol protein extraction kit and total proteins in the supernatant were extracted to perform Western blot analysis. Equal amounts of proteins were separated through electrophoresis on SDS-PAGE gels and transferred to nitrocellulose membranes. After blocked with 5% skim milk, the membranes were incubated with the appropriate primary antibodies respectively against PKC-β, and β-actin with recommended dilution at 4°C overnight. The membranes were washed and further incubated with corresponding secondary antibodies [Anti-Rabbit IgG; Anti-mouse IgG] with horseradish peroxidase (HRP) conjugation at 37 °C for 60 min. The films were developed using an ECL Plus chemiluminescence reagent kit and visualized using ChemiDocXRS. Densitometries were analyzed using Quantity One software. |
| Concentrations | 5µM |
| Incubation time | 24h |
| Animal Experiment | |
|---|---|
| Animal models | Male Sprague-Dawley rats diabetes models |
| Formulation | dissolved in citrate buffer (0.1 mol/L, pH 4.5) |
| Dosages | 1 mg/kg/day |
| Administration | oral gavage for 4 weeks |
| Molecular Weight | 505.01 |
| Formula | C28H28N4O3.HCl |
| CAS Number | 169939-93-9 |
| Solubility (25°C) | DMSO 20 mg/mL |
| Storage |
Powder -20°C 3 years ; 4°C 2 years In solvent -80°C 6 months ; -20°C 1 month |
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