Cell proliferation analyses play a pivotal role in studying cell growth and differentiation, offering insights into compound toxicity and tumor cell growth inhibition crucial for drug development. Cell proliferation assays leverage the sensitivity of fluorescent dyes to help provide accurate and reliable measurements, offering insights into cell viability, proliferation rates, and cytotoxicity.

CyQUANT cell proliferation assays are a versatile tool for researchers using a microplate format to evaluate cell growth and proliferation dynamics. Common measurements for these cell proliferation assays include DNA content or DNA synthesis. CyQUANT cell proliferation assays are particularly well-suited for high-throughput screening, boasting superior sensitivity compared to traditional colorimetric assays. These assays offer rapid and user-friendly protocols, eliminating the need for washes, extractions, or long incubations, enabling accurate results without dependence on the cell's metabolic status. Explore our array of easy-to-use CyQUANT cell proliferation assays for microplates.


CyQUANT cell proliferation selection guide

 CyQUANT Cell ProliferationCyQUANT NF (No Freeze) Cell ProliferationCyQUANT Direct Cell Proliferation AssaysCyQUANT Direct Red Cell Proliferation Assay
Use
  • Sensitive, fluorescence-based method for quantifying cells and assessing cell proliferation
  • Fast and sensitive fluorescence-based method for measuring proliferation end point assay
  • Fluorescence-based proliferation and cytotoxicity assay for microplate readers
  • Convenient no-wash workflow for high-throughput screening
  • Measurement
  • Total cell count of all live and dead cells
  • Cells must be lysed by freezing or with lysis buffer in order for stains to enter cells
  • Total cell count of all live and dead cells
  • Only live, healthy cells stained
  • Masking dye blocks staining of dead cells or cells with compromised cell membranes
  • Benefits
  • Cell samples can be frozen and stored for up to 4 weeks prior to assaying, allowing batch analyses of different time points
  • Broadest detection range
  • No lysis required for cell staining
  • Fastest incubation, 10–60 minutes
  • No lysis step allows the assay to be easily multiplexed
  • Add-mix-read protocol makes this ideal for HTS applications
  • Results correlate with metabolism-based assays
  • Linear detection range
  • 50 to 50,000 cells per well
  • Up to 250,000 with increased dye concentration
  • 100 to 20,000 cells per well
  • 50 to 20,000 cells per well
  • Incubation & assay timeMulti-day assay1 hour assay1 hour assay
    Ex/Em (nm)480/520 nm497/520 nm508/527 nm (green)622/645 nm (red)
    User guide

    Interested in measuring new DNA synthesis? Click-iT EdU Cell Proliferation Assay for microplates


    Ordering information


    Measuring DNA content

    Measuring DNA content is a crucial aspect of cell proliferation assays, and CyQUANT Cell Proliferation Assays offer an accurate microplate-based fluorescence method quantifying DNA content. Because the amount of DNA in each cell remains constant for a specific cell line or type, CyQUANT cell proliferation assays utilize this highly regulated and constant amount of cellular DNA to enable a precise measure of cell number. The CyQUANT assay can be used at multiple time points to calculate the average proliferation rate of a cell population. Binding of the CyQUANT dye to DNA is independent of metabolic state, so signal windows and fluorescence intensities can be compared across a range of conditions and cell types.

    A variety of CyQUANT assay formats provide options for different workflows, including multi-day and endpoint assays for total cell numbers and assays for live cells.

    Learn more about workflow and performance of CyQUANT cell proliferation assays below

    Measuring new DNA synthesis

    Various assays assess cell proliferation by measuring the rate of new DNA synthesis in cells. These assays involve incorporating nucleoside analogs, such as EdU or BrdU, into newly synthesized DNA. After incorporation, detection methods like click chemistry or immunostaining are used to quantify the extent of new DNA synthesis. These assays enable valuable insights into cell proliferation dynamics and can be tailored to specific experimental needs based on factors like throughput, sensitivity, and compatibility with fluorescence-based readouts.

    The Click-iT Plus EdU Microplate Assay is optimized for microplate fluorescence analysis:

    • HTS compatible—Simple and rapid workflow
    • Multiplex ability—Compatible with a range of fluorescence multiplex options
    • Signal enhancement—Amplex UltraRed dye–amplified signal (568/585, ex/em)

    Learn more about Click-iT EdU Microplate Cell Proliferation Assay

    click-it edu illustration

    Figure 1. Click-iT EdU proliferation assay for microplates

    Workflow and performance of CyQUANT cell proliferation assays


    CyQUANT cell proliferation assay

    CyQUANT Cell Proliferation Assays offer a reliable and sensitive method for quantifying cell proliferation based on cellular DNA content in a microplate format. The proprietary dye CyQUANT GR enhances fluorescence when bound to nucleic acids, allowing for the accurate measurement of cell proliferation rates by assessing DNA content within a broad linear detection range (Figure 2, 3). CyQUANT cell proliferation assays are known for their reliability in studying cell proliferation dynamics and their high sensitivity in detecting changes in cellular growth. Additionally, these assays can be easily tailored to specific experimental needs and are compatible with fluorescence-based readouts, making them versatile options for a wide range of cell proliferation studies.


    CyQUANT NF (No Freeze) cell proliferation assay

    The CyQUANT NF (no freeze) Cell Proliferation Assay offers a rapid and sensitive microplate-based method for quantifying cell proliferation without requiring cell freezing or prolonged incubations. This assay combines a cell-permeant DNA-binding dye with a membrane permeabilization reagent (Figure 4). This quantitation method uses fluorescence measurements with an excitation wavelength of 485 nm and emission detection at 530 nm, allowing for accurate determination of cell proliferation rates (Figure 5). CyQUANT NF cell proliferation assay is designed for real-time data acquisition and is well-suited for studying cell proliferation dynamics. It’s compatibility with fluorescence-based readout and the ability to customize experimental setups, CyQUANT NF assays are valuable tools for a variety of cell proliferation studies.

    CyQUANT Direct Cell Proliferation assays

    CyQUANT Direct Cell Proliferation Assays offer a straightforward and efficient method for quantifying cell proliferation and determining cytotoxicity by directly staining live cells in a microplate format without the need for wash steps or fixation. Simply grow your cells in microplate wells, add the CyQUANT direct reagent and incubate, then measure the fluorescence from the bottom of the plate. The green- and red-fluorescent CyQUANT Direct dyes stain all cells, while a background suppressor blocks staining in dead or damaged cells, ensuring only live, healthy cells are stained. CyQUANT Direct assays are highly sensitive and compatible with fluorescence-based readouts, making them suitable for studying cell proliferation dynamics in various experimental settings. With their ease of use and reliable results, these assays are valuable tools for assessing cell proliferation in research and drug discovery applications.

    The CyQUANT Direct Cell Proliferation Assay is a non-toxic, high-throughput screening method for continuous real-time monitoring of cell proliferation without cell lysis or fixation. The signal linearity and stability of this assay is a key feature that ensures reliable and accurate measurements of cell proliferation over time (Figure 6, 7). Further, the CyQUANT Direct assay is a valuable tool for simultaneously assessing cytotoxicity and cell proliferation dynamics in various experimental settings, enabling researchers to comprehensively analyze cell health and viability while evaluating the impact of compounds or experimental conditions on these responses (Figure 9).

    The CyQUANT Direct Red Cell Proliferation Assay is a variation of the CyQUANT Direct assay that incorporates a red-fluorescent DNA-binding dye, providing the same benefits with the added advantage of red fluorescence detection (Figure 6, 8, 9), making it suitable for complex experimental setups and applications requiring multiple fluorescence readouts (Figure 10).



    Figure 10. CyQUANT Direct Red assay can be multiplexed. A549 cells were seeded into the wells of a 96-well plate and incubated. After an overnight incubation, the complete medium was replaced with a staining solution containing MitoTracker Green diluted into non-serum containing medium. After a 30-minute incubation, the MitoTracker-containing medium was removed and replaced with fresh non-serum containing medium. The CyQUANT Direct Red assay was used according to manufacturer’s instructions. After a one-hour incubation, images were acquired using the EVOS FL Auto instrument equipped with the GFP (for MitoTracker Green) and the Texas Red (for detection of CyQUANT Direct Red) excitation and emission filters.


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    Resources

    Cell Analysis Learning Center
    Access educational resources for better experiment planning and execution.

    Cell Proliferation Assays Protocols
    Explore our collection of protocols for cell proliferation assays.

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    Support

    Cell Proliferation Support Center
    Find tips, troubleshooting help, and resources for your cell viability and proliferation workflow.

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    For Research Use Only. Not for use in diagnostic procedures.