Cell Proliferation ELISA, BrdU

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- Cells were seeded at a density of 1 × 104 cells/well of 96-well plates containing DMEM with 10% FBS	- Cells were treated with BrdU solution 3h before the assay

Upstream tips
- Cells were seeded at a density of 1 × 104 cells/well of 96-well plates containing DMEM with 10% FBS
Protocol tips
- Cells were treated with BrdU solution 3h before the assay

Publication protocol

BC cells (MCF-7 and MDA-MB-231) were seeded at a density of 1 × 104 cells/well of 96-well plates containing DMEM with 10% FBS. After attachments, cells were treated with different doses of ASA or DMSO for different times. Cell proliferation was determined by BrdU–ELISAs (colorimetric) according to the manufacturer’s instructions (Roche Diagnostics, Indianapolis, IN, USA). Briefly, 3 hours before the indicated time of treatment, cells were treated with BrdU solution (10 μl/well). Following BrdU treatment, cells were incubated with FixDenat solution for 30 min and then incubated with BrdU POD for 90 min at room temperature. Cells were rinsed thoroughly with washing solution and incubated with substrate solution at room temperature and processed for photometric detection.

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