Cell proliferation assay and cell cycle assay guide
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Learn about your options when you need a cell proliferation or cell cycle assay
These assays monitor the growth rate of a cell population, detect daughter cells in a growing population, or analyze the proportions of cells in different stages of the cell cycle.
Cell proliferation assay and cell cycle assay methods
- Incorporation of nucleoside analogs (eg BrdU and EdU) during DNA synthesis
- DNA staining dyes for cell cycle analysis
- Dye dilution assays
- Protein markers such as PCNA, Ki67 and MCM-2
- Clonogenicity assays
- Senescence assays
See below to learn about these types of assays. Our most popular assay kits include: EdU, propidium iodide, and CFSE.
Alternatively, cell proliferation can also be analyzed with cell viability assays that measure the rate of cellular metabolism, such as MTT, MTS, resazurin and similar assays, mitochondrial membrane potential dependent dyes, cellular esterase cleaved dyes, ATP and ADP assays, and assays that measure glycolytic flux and oxygen consumption.
Nucleoside analogs
Bromodeoxyuridine (BrdU) and ethynyldeoxyuridine (EdU) assays measure the incorporation of BrdU or EdU into newly synthesized DNA during DNA replication. Unlike BrdU, which is detected using antibodies, EdU can be easily directly labeled, either with a fluorescent dye or biotin for colorimetric or fluorometric detection via streptavidin-HRP. Edu staining is consistent with further antibody staining, unlike the harsher BrdU protocol.
EdU assay ab219801 iFluor 488 and EdU assay ab222421 iFluor 647: Microscope, flow cytometry, plate reader.
BrdU assay ab126556 and ab126572: ELISA
BrdU assay ab125306: IHC
BrdU antibody ab6326: IHC
DNA-staining dyes
DNA-staining dyes are commonly used in flow cytometry to measure the DNA content in cell populations and assay for cell cycle state. Propidium iodide is the mostly commonly used dye.
Propidium iodide ab14083 and ab139418: Ex/Em 536/617 nm
Nuclear Green CCS1 ab112116: Ex/Em 490/525 nm
Nuclear Red CCS1 ab112117: Ex/Em 490/620 nm
DRAQ5™ ab108410: Ex/Em 633 & 647/665–800 nm
DAPI ab228549: Ex/Em 358/461
Hoechst 33342 ab228551: Ex/Em 350/461
Hoechst 33258 ab228550: Ex/Em 350/461
7-AAD ab228563: Ex/Em 488/647 nm
The dyes in dye dilution assays are retained within cells over multiple generations. Daughter cells receive half of the dye of parent cells and assays are analyzed on a flow cytometer. Carboxyfluorescein succinimidyl ester (CFSE) is the longest established dye.
CFSE assay ab113853: Ex/Em 492/517 nm. Cytotoxic at higher concentrations. Flow cytometer.
CytoLabel Blue ab176726: Ex/Em 403/454 nm. Flow cytometer, microscope.
CytoLabel Green ab176735: Ex/Em 511/525 nm. Flow cytometer, microscope.
CytoLabel Red ab176736: Ex/Em 628/643 nm. Flow cytometer, microscope.
CytoLabel Orange ab176737: Ex/Em 542/556 nm. Flow cytometer, microscope.
For the analysis of cell proliferation within tissue samples, or sometimes within cell culture, it is common to use antibodies to stain for the presence of Ki67, PCNA, or MCM-2.
Although little used at high throughput, the classical method of assaying cell proliferation is to use a clonogenic/clonogenicity assay. In this assay, cells are plated out at a low density and then the number of colonies formed is counted.
The most common marker of senescent cells is the overexpression and accumulation of the endogenous lysosomal beta-galactosidase (SA-beta-gal). Beta-gal activity is detected using a colorimetric or fluorometric substrate.
Beta-galactosidase assay ab65351: Microscope, plate reader.
Learn more with our:
Cell viability assay guide
Measure the rate of continuing cellular activities, such as metabolism.
Cytotoxicity assay guide
Test for cell membrane damage, either by measuring the leakage of cellular enzymes or staining with membrane-impermeable dyes.
Apoptosis assay / cell death analysis guide
Measure the markers present in different types of cell death.