Western blot analysis of extracts from C2C12 cells, untreated (-) or treated with Forskolin #3828 (10 μM, 30 min; +) with or without IBMX pretreatment (4 hr) at the indicated concentrations, using Phospho-VASP (Ser157) Antibody #3111 (upper), Phospho-VASP (Ser239) Antibody #3114 (middle), or VASP (9A2) Rabbit mAb #3132 (lower).
Chemical structure of IBMX.
IBMX is supplied as a lyophilized powder. For a 250 mM stock, reconstitute the 50 mg in 899.7 µl DMSO. Working concentrations and length of treatment can vary depending on the desired effect, but it is typically used at 10-1,000 µM for 15 min-4 hr.
Store lyophilized or in solution at -20ºC, desiccated. 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.
3-isobutyl-1-methylxanthine (IBMX) is a nonselective inhibitor of cAMP and cGMP phosphodiesterases (PDEs). Research studies show that IBMX inhibits many members of the PDE family, with the exceptions of PDE8A, PDE8B, and PDE9 (1-4). Treatment of cells with IBMX promotes accumulation of cAMP and cGMP, which leads to activation of cyclic-nucleotide-regulated protein kinases (5,6). Additional research indicates that IBMX can promote neuronal progenitor cell maturation in vitro (7).
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