Cell Proliferation Assays for Flow Cytometry

Click-iT Edu assays for flow cytometry

Measure DNA synthesis with Click-iT flow cytometry kits. The kits are optimized for live cells, the click detection step comes after the fixation.

Protocol: Click-iT EdU flow cytometry cell proliferation assays

A common method to measure DNA synthesis involves the incorporation of BrdU into newly synthesized DNA in living cells, which can later be detected using a fluorescently labeled antibody that recognizes BrdU. The fluorescent signal can be analyzed with flow cytometry instruments to provide a quantitative measurement of DNA synthesis during cell proliferation. 

Click-iT EdU flow cytometry assay kits were developed to overcome some of limitations of the BrdU technique. Click-iT EdU and Plus EdU flow cytometry assays utilize the power of click chemistry and the modified nucleoside EdU to offer an excellent alternative to BrdU staining for detecting and quantitating newly synthesized DNA.

• Accuracy—typically better than BrdU assays with minimal variation (low CVs)
• Simplicity—streamlined protocol
• Compatibility—assays allow users to multiplex with GFP, mCherry, APC, PerCP, PE, and other fluorophores (Click-iT Plus EdU kits only)

Explore: Click-iT EdU cell proliferation assays

CellTrace stains are fluorescent tracking dyes that are retained in living cells. With each generation, the amount of dye in each cell is halved, so fluorescence analysis of proliferating populations can be used to quantify the successively decreasing brightness for each generation. These dyes are typically effective at reporting proliferation above background autofluorescence for over six generations.

• Excellent performance—bright, single-peak staining enables visualization of multiple generations
• Long-term signal stability—well retained in cells for several days post-stain
• Non-cytotoxic—no known effect on proliferative ability or biology of cells
• Compatibility—bright, single-peak staining available in multiple fluorophores
• Versatile—multiple colors available to easily combine with antibodies or markers of cell function, such as GFP

Analysis of the level of fluorescence in the cell populations by flow cytometry permits the determination of the number of generations through which a cell has progressed since the label was applied (Figure 1).

Cell cycle phases coordinate multiple proteins to synthesize DNA and divide cells. Building flow cytometry panels for the detection of cell proliferation markers can help evaluate this process. Common cell proliferation markers include 5-bromodeoxyuridine (BrdU), Ki-67, minichromosome maintenance (MCM2), and proliferating cell nuclear antigen (PCNA).