Chemical structure of PKC412.
Western blot analysis of extracts from Jurkat cells, untreated or treated with Calyculin A #9902 (50 nM, 20 min) with or without pre-treatment with PKC412 (24 hr) for the indicated concentrations, using Phospho-(Ser) PKC Substrate Antibody #2261 (upper) and β-Actin (13E5) Rabbit mAb #4970 (lower).
Western blot analysis of extracts from SEM cells, untreated (-) or treated with PKC412 (1 μM, 24 hr; +), using Phospho-FLT3 (Tyr589/591) (30D4) Rabbit mAb #3464 (upper) and FLT3 (8F2) Rabbit mAb #3462 (lower).
PKC412 is supplied as a lyophilized powder. For a 10 mM stock, reconstitute 1 mg in 175.24 μL DMSO. Working concentrations and length of treatment can vary depending on the desired effect, but it is typically used at 0.1-10 μM for 18-48 hours. Soluble in DMSO at 100 mg/mL; soluble in ethanol at 5 mg/mL with warming; very poorly soluble in water with maximum solubility estimated to be about 10-20 μM.
Store lyophilized or in solution at -20ºC, desiccated. Protect from light. In lyophilized form, the chemical is stable for 24 months. Once in solution, use within 3 months to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.
The staurosporine analog, PKC412, inhibits a variety of serine/threonine and tyrosine kinases including PKC (IC50 = 50 nM), cyclic AMP-dependent protein kinase (IC50 = 2.4 µM), S6 kinase (IC50 = 5.0 µM), and EGFR (epidermal growth factor receptor) tyrosine kinase activity (IC50 = 3.0 µM) (1). PKC412 has also demonstrated stong inhibitory effects on FLT3, which causes G1 cell cycle arrest and apoptosis (2).
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