Cell proliferation assays and cell cycle analysis
Cell proliferation assays are vital in biological and medical research for evaluating the growth and division rates of cell populations, offering insights into cellular behaviors crucial for understanding biological processes, disease mechanisms, and therapeutic outcomes. These assays monitor cell multiplication, quantify new daughter cells, or assess cell cycle distribution, essential for studying cancer progression, tissue regeneration, drug efficacy, and immune response.
Common methods for assessing cell proliferation include:
- Cell proliferation marker antibodies: These antibodies image proteins indicating cell division, providing visual insights within tissue samples.
- BrdU and EdU assays: These highly sensitive assays detect DNA synthesis in actively dividing cells.
- Metabolic assays: By measuring metabolic activity, researchers can infer the number of viable cells based on substrate processing through enzymatic pathways.
- Fluorescent dye-based assays: These involve staining cells with fluorescent dyes to analyze their cycle stage or division rate, offering a precise and quantitative approach.
Cell proliferation assessment is crucial in cancer biology for informing treatment strategies, in drug development for evaluating compound cytotoxicity, and in regenerative medicine for tracking stem cell differentiation and tissue repair. Thus, cell proliferation assays are indispensable for advancing scientific discovery and clinical applications.
Researchers commonly assess cytotoxicity, mitochondrial function, and caspase activity in addition to proliferation. For a deeper dive into cell health, including the background on methods to assess cell viability, cytotoxicity, mitochondrial function, the roles of the various caspases, and more, download our Cell Health Guide.
To view our range of products, including recommended products, for assessing apoptosis, view our apoptosis and apoptotic markers product guide.
Cell proliferation markers for imaging
Antibodies targeting proliferation markers detect specific proteins expressed exclusively or at higher levels during certain cell cycle stages. These markers, used in imaging and flow cytometry, quantify cell division in tissue sections and cell cultures through chromogenic or fluorescent labels, offering application versatility. Key markers include Ki67, PCNA, and MCM2:
Ki67, present during all active cell cycle phases (G1, S, G2, M) but absent in resting cells (G0), reliably indicates cell proliferation across various tissues and conditions.
PCNA, crucial for DNA replication during S-phase and DNA repair, allows detection of cells undergoing DNA synthesis and repair, aiding studies on cancer, tissue regeneration, and drug efficacy.
MCM2, part of the helicase complex essential for DNA replication initiation, serves as a precise marker for cells entering S-phase.
Abcam provides a comprehensive range of antibodies against these and other proliferation markers, validated for human, mouse, rat, and more. Suitable for paraffin-embedded or frozen tissues and cell cultures, our highly cited antibodies underscore their reliability and performance in diverse research applications.
Product highlights
Anti-Ki67 antibody [SP6]
- Cited in >2,500 scientific publications
- Can be used for IHC, immunofluorescence, western blotting and flow cytometry
- Can be used on human, mouse, and rat samples
Anti-PCNA antibody [EPR3821]
- Can be used for IHC, immunofluorescence, western blotting, immunoprecipitation, and flow cytometry
- Can be used on human, mouse, and rat samples
Anti-MCM2 antibody [EPR4120]
- Can be used for IHC, immunofluorescence, western blotting, immunoprecipitation, and flow cytometry
- Can be used on human, mouse, and rat samples
Annexin V/ANXA5-FITC Apoptosis Detection Reagent (500X)
- Annexin V FITC conjugate used in ab14085
DNA synthesis assays for cell proliferation
DNA synthesis in a sample directly correlates with cell proliferation, as dividing cells replicate their DNA. This can be indirectly assessed using DNA-staining fluorescent dyes like propidium iodide (PI) in cell cycle assays. For direct measurement, synthetic nucleoside analogs are incorporated into cells or tissues, reflecting DNA synthesis during replication.
The BrdU (Bromodeoxyuridine) and EdU (ethynyldeoxyuridine) assays are commonly used for measuring DNA synthesis. Both involve the incorporation of these analogs into replicating DNA, allowing precise quantification. BrdU is detected using antibodies, suitable for various applications but time-consuming due to the necessary denaturation step for antibody binding. Conversely, EdU allows simpler and more efficient detection, directly labeled with fluorescent dyes or biotin, eliminating harsh DNA denaturation steps and making it faster and more user-friendly.
Both BrdU and EdU offer safer and more practical alternatives to older methods using radioactive nucleotides like tritiated thymidine, which pose health and environmental risks. Kits are available for purifying fragmented DNA from cultured cells, facilitating easy analysis of DNA synthesis. Purified DNA fragments can be run on an agarose gel for further examination, providing a straightforward approach to assessing DNA replication in research.
We supply the gold-standard BrdU antibody clone BU1/75 (ICR1), which has been used in thousands of publications and is used for BrdU imaging in cells and tissue sections.
We also supply a BrdU ELISA kit, which can quantify the amount of BrdU incorporated into cells in a sample, and several EdU staining kits for fluorescent imaging, offering a shorter staining protocol than that used with BrdU staining.
Product highlights
Anti-BrdU antibody [BU1/75 (ICR1)] - Proliferation Marker
- Gold standard BrdU antibody
- Used in thousands of publications
- Suitable for imaging and flow cytometry
BrdU Cell Proliferation ELISA Kit (colorimetric)
- Convenient, complete kit for staining cells with BrdU
- Allows for quantitative detection by ELISA
EdU Assay / EdU Staining Proliferation Kit (iFluor 488)
- EdU labeling kit for flow cytometry and imaging
- Faster assay with fewer steps than using BrdU
Metabolic assays for cell proliferation
Metabolic assays for cell proliferation rely on living cells metabolizing specific chemicals via enzymes, which can be co-opted to convert non-fluorescent compounds into fluorescent or colorimetric products. The signal intensity correlates with the number of living cells, providing a reliable method to assess viability and proliferation. These non-invasive assays enable real-time monitoring of cell cultures. Another method involves lysing cells and measuring ATP levels, which indicate cellular energy and viability, correlating with live cell counts. This approach is useful in cytotoxicity testing and drug efficacy studies.
Abcam offers various ready-to-use assay kits for metabolism-based cell proliferation assays, facilitating accurate and rapid data collection in diverse experimental contexts, whether through enzymatic reactions or ATP quantification.
Product highlights
MTT Assay Kit (Cell Proliferation)
- Easy-to-use
- Measure cell proliferation, cell viability, and cytotoxicity
- High-throughput assay for
MTS Assay Kit (Cell Proliferation) (Colorimetric)
- Allows for sensitive quantification of viable cells
MTS Assay Kit (Cell Proliferation) (Colorimetric)
- Easy-to-use
ATP Assay Kit (Colorimetric/Fluorometric)
- Mix-and-read protocol
- No-wash assay
- Only one 30 min incubation
Fluorescent dye-based cell cycle assays
DNA-staining fluorescent dyes, like propidium iodide (PI), are extensively used in flow cytometry to evaluate DNA content and identify the cell cycle stages, distinguishing between G0/G1, S, and G2/M phases. An alternative method, the dye dilution assay, involves labeling cells with a fluorescent dye retained over generations. As cells divide, the dye is equally distributed between daughter cells, allowing the tracking of cell divisions through fluorescence dilution via flow cytometry. Carboxyfluorescein succinimidyl ester (CFSE) is a prime dye for this method due to its long-lasting fluorescence, ideal for monitoring extended cell proliferation. Both PI and CFSE are invaluable for analyzing cell cycle dynamics and division history, crucial in fields like cancer biology, immunology, and drug development. These assays are popular in flow cytometry for their reliability and ease in tracking cell proliferation and cycle progression.
Product highlights
Propidium Iodide Flow Cytometry Kit
- Can be used with both cells in cells grown in suspension and adherent cells
CFSE - Cell Labeling Kit
- Non-toxic
- Uniform bright fluorescence signal
