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measuring cell proliferation with SpectraMax M5, M2 and Gemini microplate readers using the CyQUANT Cell Proliferation Assay

In the first, cellular proliferation is quantitated using a cell-based standard curve. In the second, cellular proliferation is quantitated using RNase-treated cell samples and a DNA standard curve.


MATERIALS

  • CyQUANT Cell Proliferation Assay Kit (Molecular Probes Cat. # C-7026): Component A, CyQUANT GR dye at 400X; component B, cell-lysis buffer at 20X; and component C, l DNA standard, 100 L of a 100 g/mL solution

  • Note: Lysis Buffer and CyQUANT GR should be used within a few hours of being diluted. Be sure to protect the CyQUANT GR solution from light.

  • Cells: CHO-K1 cells were used to generate the data in this application note (ATCC # CCL-61)

  • EDTA if using a DNA standard curve (Fisher Chemicals Cat. # S311-100)

  • NaCl if using a DNA standard curve (Fisher Chemicals Cat. # S271-500)

  • DNase-free RNase A or RNase cocktail (Ambion Cat. # 2286)

  • Black 96-well clear-bottom microplates (Corning Cat. # 3603)

  • SpectraMax M2/M5 Multi-Detection Microplate Readers (Molecular Devices)

  • Gemini XPS/EM Microplate Fluorometers (Molecular Devices)


    CELL PREPARATION
    Adherent or non-adherent cells were prepared for the CyQUANT assay by growing and freezing them directly in the microplate(s) in which they were to be assayed, or by growing them in culture vessels, then freezing pelleted samples of the cells to be later transferre
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