Western blot analysis of extracts from NIH/3T3 cells, untreated (-) or treated (+) with Bortezomib (100 nM, 6 hr), using Ubiquitin Antibody #3933 (left), p53 (1C12) Mouse mAb #2524 (middle), and β-Actin (13E5) Rabbit mAb #4970 (right).
Chemical structure of Bortezomib.
Western blot analysis of L363 cells, untreated (-) or treated (+) with Bortezomib (24 hr) as indicated, using K48-linkage Specific Polyubiquitin Antibody #4289.
Bortezomib is supplied as a lyophilized powder. For a 1 mM stock, reconstitute the 2.5 mg in 6.51 ml DMSO. Working concentrations and length of treatment can vary depending on the desired effect, but it is typically used at 1-1000 nM for 2-48 hours. Soluble in DMSO and ethanol at 200 mg/ml, very poorly soluble in plain water.
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.
Bortezomib (C19H25BN4O4), a boronic acid dipeptide, is a specific, potent, and reversable proteasome inhibitor that has also been shown to have a stabilizing effect on proteins that inhibit cell survival and cell cycle progression, such as p53 (1,2). Researchers have demostrated that bortezomib inhibits activation and nuclear translocation of NF-κB, subsequently decreasing early tumor survival (3). The increase in bortezomib-stablilized, misfolded, and ubiquitinated proteins that interfere with cell survival and other important pathways further support its anti-tumor effects (3).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.