Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Mitomycin C (2 μg/mL, 24 hr; +), using FANCD2 (D5L5X) Rabbit mAb #16323 (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). DNA damage caused by Mitomycin C induces monoubiquitination of FANCD2, altering its electrophoretic mobility and increasing its apparent molecular weight (5).
Chemical structure of Mitomycin C.
Mitomycin C is supplied as a lyophilized powder. For a 15 mM stock, reconstitute 5 mg of powder in 997 μl of DMSO. Working concentrations and length of treatment can vary depending on the desired effect.
Store lyophilized at -20ºC, desiccated. In lyophilized form, the chemical is stable for 24 months. Once in solution, store at -20ºC and use within 1 month to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.
Molecular Weight: 334.3 g/mol
Molecular Formula: C15H18N4O5
Solubility: Soluble in DMSO at 15 mg/ml or water at 1 mg/ml with slight warming.
Mitomycin C, also known as MMC, is an antitumor antibiotic isolated from Streptomyces caespitosus (1). When activated, this alkylating agent crosslinks double stranded DNA resulting in DNA damage (2). Mitomycin C has been found to have lethal effects on HeLa cells during the G1 phase of the cell cycle (3). DNA damage caused by Mitomycin C can lead to increased levels of p53 that can result in cell cycle arrest or apoptosis (4).
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