BrdU Cell Proliferation Assay

Cell cytotoxicity / Proliferation assay cell type - Panc-1

Experiment
Cell cytotoxicity / Proliferation assay cell type - Panc-1
Product
BrdU Cell Proliferation Assay from Millipore
Manufacturer
Millipore

Protocol tips

Upstream tips
- 3×103 cells/well in a 96-well plate were seeded for an assay.
Protocol tips
- After 24 h or culture, for another 2h cells were incubated with BrdU.

- anti-BrdU monoclonal antibody isincubated for 1 hr.

Publication protocol

Cell proliferation was also assessed by 5′-bromo-2′-deoxyuridine (BrdU) incorporation. The BrdU Assay kit (Millipore) was used according to the manufacturer’s recommendations. Cells were seeded at a density of 3×103 cells/well in a 96-well plate. Following 24 hr of culture, BrdU was added and allowed to accrue within proliferating cells for 2 hr. The cells were then fixed at room temperature for 30 min followed by incubation with an anti-BrdU monoclonal antibody for 1 hr. The cells were then incubated at room temperature with a secondary antibody conjugated with peroxidase. The substrate solution tetramethylbenzindine (TMB) was added with incubation for 30 min in the dark at room temperature. The staining reaction was terminated with a 2.5 N sulfuric acid solution and the plate read with a spectrophotometer at 450 nm.

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Papers

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Paper title
IL-6 stimulates STAT3 and Pim-1 kinase in pancreatic cancer cell lines
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Manufacturer protocol

Download the product protocol from Millipore for BrdU Cell Proliferation Assay below.

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