Cell Death Detection ELISA

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	Cell apoptosis was detected using a photometric enzyme immunoassay (Cell Death Detection ELISA kit, cat. no. 11544675001; Sigma-Aldrich Merck KGaA), as previously described (30). This is based on the quantitative sandwich immunoassay employing antibodies against histone and DNA. MDA-MB-231 cells and MCF-7 cells (1×105 cells/well) were seeded into a 6-well plate in the presence or absence of different concentrations of BBM (10, 20 and 40 µM) and cultured for 24 h at 37°C. The cells were washed with PBS and then lysed in RIPA lysis buffer (containing 1 mM PMSF). The supernatant was collected via centrifugation at 12,000 × g for 15 min at 4°C. and used for testing. The mono- and oligonucleosomal fragmented DNA was measured according to the instructions of the manufacturer.

Protocol tips

Cell apoptosis was detected using a photometric enzyme immunoassay (Cell Death Detection ELISA kit, cat. no. 11544675001; Sigma-Aldrich Merck KGaA), as previously described (30). This is based on the quantitative sandwich immunoassay employing antibodies against histone and DNA. MDA-MB-231 cells and MCF-7 cells (1×105 cells/well) were seeded into a 6-well plate in the presence or absence of different concentrations of BBM (10, 20 and 40 µM) and cultured for 24 h at 37°C. The cells were washed with PBS and then lysed in RIPA lysis buffer (containing 1 mM PMSF). The supernatant was collected via centrifugation at 12,000 × g for 15 min at 4°C. and used for testing. The mono- and oligonucleosomal fragmented DNA was measured according to the instructions of the manufacturer.

Publication protocol

Cell apoptosis was detected using a photometric enzyme immunoassay (Cell Death Detection ELISA kit, cat. no. 11544675001; Sigma-Aldrich Merck KGaA), as previously described (30). This is based on the quantitative sandwich immunoassay employing antibodies against histone and DNA. MDA-MB-231 cells and MCF-7 cells (1×105 cells/well) were seeded into a 6-well plate in the presence or absence of different concentrations of BBM (10, 20 and 40 µM) and cultured for 24 h at 37°C. The cells were washed with PBS and then lysed in RIPA lysis buffer (containing 1 mM PMSF). The supernatant was collected via centrifugation at 12,000 × g for 15 min at 4°C. and used for testing. The mono- and oligonucleosomal fragmented DNA was measured according to the instructions of the manufacturer.

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