DELFIA Cell Proliferation Assay

Figure 2. Workflow for DELFIA cell proliferation assay

Citations

View a brief list of DELFIA cell proliferation citations.

Using the DELFIA cell proliferation kit with suspension cells

1. Place appropriate number of cells in a 96-well plate (e.g., in a final volume of 200 µL per well) and incubate them with the substance to be tested at 37°C in a humidified 5% CO2 atmosphere. The incubation period depends on the cell type used. For most experimental approaches, an incubation period of 24-120 hours is appropriate.
2. Label cells with BrdU by adding 20 µL of diluted (100 µM) BrdU Labeling solution to each well if the cells were cultured in 200 µL of culture medium. Continue incubating the cells for an additional 2 to 24 hours at 37°C in a humidified 5% CO2 atmosphere. NOTE: The volume of BrdU Labeling solution to be added depends on the volume of the cell culture, and the final concentration of BrdU in the wells should be 10 µM.
3. Centrifuge the plate at 300 x g for 10 minutes before removing the labeling medium. After removing the medium, incubate at 60°C for 1 hour or until the wells are dry.
4. Add 100 µL of Fix solution to each well and incubate for 30 minutes at room temperature.
5. Remove the Fix solution thoroughly from the wells either by inverting the plate and shaking it, or by aspiration.
6. Add 100 µL of anti-BrdU-Eu working solution (0.5 µg/mL) to each well and incubate for 30--120 minutes at room temperature.
7. Wash 4 times using the DELFIA Platewash with approximately 300 µL of Wash solution per well.
8. Add 200 µL DELFIA Inducer to each well using the DELFIA Plate Dispense or Eppendorf Multipipette. When using the DELFIA Plate Dispense, make sure that the tubing is flush with DELFIA Inducer before dispensing into the well. Shake the plate on the DELFIA Plateshake at room temperature for 15 minutes (slow shaking). When using a different shaker, check that the liquid is moving constantly.
9. Measure the Eu-fluorescence in a time-resolved fluorometer, such as VICTOR™2V or the EnVision™ using parameters recommended in the kit manual.

For research use only. Not for use in diagnostic procedures.  

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