Western blot analysis of extracts from HeLa cells serum-starved overnight, and untreated (-) or treated with PDBu (15 min; +) at the indicated concentrations, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (upper) or p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).Learn more about how we get our images
Chemical structure of PDBu.Learn more about how we get our images
PDBu is supplied as a lyophilized powder. For a 2 mM stock, reconstitute the 1 mg in 990.9 µl DMSO. Working concentrations and length of treatment can vary depending on the desired effect, but it is typically used at 20-2000 nM for 15-30 min. Soluble in DMSO or ethanol.
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.
Phorbol 12, 13-Dibutyrate (PDBu) is a phorbol ester and a potent activator of protein kinase C (PKC). Frequently used in cancer research, PDBu is widely characterized to promote tumors in a variety of tissue types (1,2). PDBu treatment of cells can lead to activation of downstream PKC targets such as p44/42 MAPK and MARCKS (3-5). Although less potent than TPA, research has shown PDBu elicits less nonspecific binding (6,7) and is less hydrophobic allowing for easy wash out from cells in tissue culture (8).
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