All tags Cell cycle Cell proliferation assay and cell cycle assay guide

Cell health

Cell proliferation assay and cell cycle assay guide

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

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.

Assay

Instrument

Assay kits

EdU

Microscope, flow cytometry, plate reader

ab219801
ab222421

BrdU

Plate reader, microscope, flow cytometry

ELISA: ab126556 / ab126572    
IHC: ab125306
Antibody: ab6326


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.

Dye

Instrument

Note

Assay kits

Propidium iodide

Flow cytometer

Ex/Em 536/617 nm

ab14083, ab139418

Nuclear Green CCS1

Ex/Em 490/525 nm

ab112116

Nuclear Red CCS1

Ex/Em 490/620 nm

ab112117

DRAQ5TM

Ex/Em 633 & 647/665–800 nm

ab108410

DAPI

Ex/Em 358/461


Hoechst 33342

Ex/Em 350/461


Hoechst 33258

Ex/Em 350/461


7-AAD 

Ex/Em 488/647 nm


Dye dilution assays

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.

Assay

Instrument

Notes

Assay kits

CFSE

Flow cytometer

Ex/Em 492/517 nm. Cytotoxic at higher concentrations.

ab113853

CytoLabel Blue

Flow cytometer, microscope

Ex/Em 403/454 nm

ab176726

CytoLabel Green

Ex/Em 511/525 nm

ab176735

CytoLabel Red

Ex/Em 628/643 nm

ab176736

CytoLabel Orange

Ex/Em 542/556 nm

ab176737

Protein markers

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.


Clonogenicity assays

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.

Senescence assays

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.

Assay

Instrument

Assay kits

Beta-gal

microscope, plate reader

ab65351


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.



注册