Application Notes
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Optimal working dilution should be determined by the investigator.
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Comment
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- Contains sufficient reagents for the evaluation of 960 assays in ten 96-well plates or 192 assays in eight 24-well plates
- Can be applied to most eukaryotic cell lines, including adherent and non-adherent cells and certain tissues
- Cell proliferation reagent can be used to detect proliferation in bacteria, yeast, fungi, protozoa as well as cultured mammalian and piscine cells
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Protocol
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CytoSelect? Cell Proliferation Assay Reagent (Fluorometric) provides a fluorometric format for measuring and monitoring cell proliferation. The kit contains sufficient reagents for the evaluation of 960 assays in ten 96-well plates or 192 assays in eight 24-well plates. Cells can be plated and then treated with compounds or agents that affect proliferation. Cells are then incubated with the proliferation reagent. Upon entering metabolically active live cells, the non-fluorescent proliferation reagent is converted into a bright red fluorescent form. An increase in cell proliferation is accompanied by increased fluorescent signal, while a decrease in cell proliferation (and signal) can indicate the toxic effects of compounds or suboptimal culture conditions. The assay principles are basic and can be applied to most eukaryotic cell lines, including adherent and non-adherent cells and certain tissues. This cell proliferation reagent can be used to detect proliferation in bacteria, yeast, fungi, protozoa as well as cultured mammalian and piscine cells.
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Assay Procedure
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- Prepare a cell suspension containing 0.1-1.0 x 106 cells/mL in medium.
- Add 100 μL of cell suspension per well to a 96-well cell culture plate or 500 μL per well to a 24- well cell culture plate with or without the compound to be tested. Culture the cells for 24-96 hours at 37?°C and 5?% CO2 in a humidified incubator.
- Add one tenth volume of the CytoSelect? Cell Proliferation Assay Reagent per well (for example add 10 μL of CytoSelect? Cell Proliferation Assay Reagent per 100 μL of cell culture).
- Incubate plate at 37?°C and 5?% CO2 for 4-8 hours.
- Read the fluorescence with a fluorescence plate reader at 560 nm excitation wavelength and 590- 600 nm emission wavelength.
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Restrictions
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For Research Use only
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