Cell proliferation analyses are crucial for cell growth and differentiation studies, and are often used to evaluate both compound toxicity and inhibition of tumor cell growth during drug development. Proliferation measurements are typically made based on average DNA content or on cellular metabolism parameters. Assays can report either total or live cell numbers, or measure DNA synthesis in single cells.
Molecular Probes® cell proliferation assays can be used individually or together to monitor cell health and cell division by evaluating DNA synthesis, monitoring mitotic phases, or tracking population doubling.
Measuring the synthesis of new DNA is a precise way to assay cell proliferation in individual cells or in cell populations. DNA synthesis–based cell proliferation assays measure the rate of new DNA synthesis based on incorporation of modified nucleosides. The Click-iT® Plus EdU cell proliferation assay utilizes the power of click chemistry and the modified nucleoside EdU to provide a superior alternative to BrdU staining for detecting and quantitating newly synthesized DNA.
Measuring cell proliferation with Click-iT® Plus EdU assays also means that cell proliferation measurements can be easily multiplexed with expressed proteins like GFP, protein labels like R-PE, and a wide range of organic fluorophores.
- Accuracy—superior to BrdU assays with minimal variation (low CVs)
- Simplicity—streamlined five-step protocol
- Compatibility—multiplex with GFP, mCherry, APC, PerCP, PE, and other fluorophores
Learn more about our DNA synthesis measurement assays for flow cytometry ›
Dye dilution assays for cell proliferation rely on cell membrane–permeant fluorescent molecules. Upon entry into the cell, the dye will covalently bind to amine groups on proteins, resulting in long-term dye retention within the cell.
Through subsequent cell divisions, each daughter cell receives approximately half the fluorescence of the parent. Analysis of the fluorescence intensities of cell populations by flow cytometry enables determination of the number of generations through which a cell or population has progressed since the label was applied.
CellTrace™ fluorescent stains can be used without affecting morphology or physiology to trace generations in vivo or in vitro.
- Long-term signal stability—well-retained in cells for several days post-stain
- Noncytotoxic—no known effect on proliferative ability or biology of cells
Learn more about dye dilution assays for flow cytometry
|Click-iT® Plus EdU Pacific Blue™ Flow Cytometry Kit||Click-iT® Plus EdU Alexa Fluor® 488 Flow Cytometry Kit||Click-iT® Plus EdU Alexa Fluor® 647 Flow Cytometry Kit|
|Basis of assay||A replacement for traditional BrdU assays, the modified thymidine analog EdU is incorporated into newly synthesized DNA in proliferating cells. It labels the cells with a bright, photostable Alexa Fluor® dye in a fast, highly specific click reaction.|
|Readout||Fluorescent signal accumulates in the nuclei of cells where DNA has been synthesized during the EdU incubation period.|
|Fluoresent label||Pacific Blue® picolyl azide||Alexa Fluor® 488 picolyl azide||Alexa Fluor® 647 picolyl azide|
|Multiplexable||The mild click chemistry protocol does not destroy cellular epitopes (unlike BrdU protocols) and is compatible with antibody labeling, fluorescent proteins, and common cell staining methods.|
|Sample type||Optimized for live cells, the click detection step comes after the fixation.|
|Format||50 assays||50 assays||100 assays||50 assays||100 assays|
|CellTrace™ Violet Cell Proliferation Kit, for Flow cytometry||CellTrace™ CFSE Cell Proliferation Kit - For Flow Cytometry||CellTrace™ Far Red DDAO-SE Proliferation Kit, for Flow Cytometry|
|Basis of assay||Permanently label cells with fluorescent stains to trace generations or divisions in-vivo or in-vitro without affecting morphology or physiology.|
|Readout||The level of fluorescence in the cell populations determines the number of generations which a cell has progressed since the fluorescent label was applied.|
|Fluoresent label||CellTrace™ Violet||CFSE||DDAO-SE|
|Format||1 kit/180 tests||1 kit||20 x 50 µg|
Cell proliferation scientific posters