Western blot analysis of extracts from HeLa cells, serum-starved overnight and untreated or treated with Doxorubicin (5 μM) for the indicated times, using Phospho-p53 (Ser15) Antibody #9284, p53 Antibody #9282, Cleaved Caspase-3 (Asp175) Antibody #9661, or PARP Antibody #9542.
Western blot analysis of extracts from HeLa cells, untreated, treated with Doxorubicin (0.5 µM, 24 hr), or treated with Trichostatin A (TSA) #9950 (400 nM, 24 hr) and Doxorubicin (0.5 µM, 24 hr), using Acetyl-p53 (Lys382) Antibody #2525 (upper) or p53 (1C12) Mouse mAb #2524 (lower).
Chemical structure of doxorubicin.
Doxorubicin is supplied as a lyophilized powder. For a 10 mM stock, reconstitute the 5 mg in 860 µl DMSO. Working concentrations and length of treatments vary depending on the desired effect, but it is typically used at 0.1-5 µM for 12-24 hours. Soluble in DMSO at 100 mg/ml; very poorly soluble in ethanol; soluble in water at 10 mg/ml with slight warming.
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.
C27H29NO11 • HCl
Doxorubicin, an anthracycline antibiotic, inhibits DNA and RNA synthesis in mammalian cells and has been shown to be a very effective anti-tumor agent (1,2). Doxorubicin binds to nucleic acids by intercalating the DNA double helix and stabilizing topoisomerase II cleavage complexes, leading to DNA strand breaks at specific doxorubicin-induced sites (3). Doxorubin has been shown to inhibit DNA synthesis in a dose-dependent manner in MCF7 cells, which corresponds closely with growth inhibition (4). Researchers have also demonstrated that doxorubicin effectively inhibits human DNA topoisomerase I (5).
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