EdU T Cell Proliferation Assay for Flow Cytometry

High-resolution detection of T-Cell Proliferation with ClickTech EdU Kit

The ClickTech EdU T-cell Proliferation Kit is a powerful tool for the direct detection of DNA synthesis in activated human T cells via click chemistry-based EdU incorporation. Unlike dye-dilution methods, this kit precisely detects cells actively synthesizing DNA during S-phase of the cell cycle, providing high-resolution data even in complex co-culture or immunotherapy-relevant systems.

Figure 1: Flow cytometry histogram showing T cell proliferation detected via EdU incorporation and 6-FAM azide staining. The histogram displays two distinct populations:

Proliferating T cells (EdU⁺) are represented by the larger peak at higher fluorescence intensity (colored in light green), indicating successful incorporation of EdU during DNA synthesis.

Non-proliferating T cells (EdU⁻) form a separate peak at lower fluorescence intensity (gray), reflecting cells that did not enter S-phase during the EdU pulse.

Key Features

• Direct detection of proliferating T cells through EdU incorporation
• Ideal for immunotherapy, vaccine research, T-cell manufacturing, and drug screening
• No need for harsh DNA denaturation steps (unlike BrdU)
• Multiplex with surface and intracellular markers for in-depth immune profiling

Applications

• Monitoring T-cell activation and proliferation post-stimulation (e.g., CD3/CD28)
• Characterizing antigen-specific T-cell responses
• Assessing proliferation in chimeric antigen receptor T-cell therapy (CAR-T) and checkpoint inhibitor studies
• Evaluating donor–recipient responses in Mixed Lymphocyte Reactions (MLR)
• Tracking T-cell expansion in bioreactors and ex vivo cultures
• High-content immunophenotyping with transcription factors or cytokines

Why EdU instead of dye dilution?

Dye dilution methods like CellTrace™ Far Red or CFSE are commonly used to track T-cell divisions. However, they can suffer from:
• Inconsistent staining in complex populations
• Signal loss over time, complicating long-term studies
• Interference with cell function or downstream staining

The ClickTech EdU Kit offers a reliable, DNA-level readout of cell proliferation unaffected by cell type, metabolic state, or dye efflux.

Compare with other proliferation assays

Kit components

• EdU reagent (optimized for T-cell incorporation)
• ClickTech detection buffer system
• Permeabilization & fixation buffers
• Detailed protocol optimized for human T-cell activation workflows

Workflow overview

• Stimulate T cells (e.g., anti-CD3/CD28, antigens, APCs)
• Add EdU to culture medium
• Incubate (typically 4–24 hours)
• Harvest and stain cells using our optimized click chemistry protocol
• Analyze via flow cytometry (can be multiplexed with CD3, CD4, CD8, CD25, FoxP3, IFN-γ, etc.)

Available Formats

• 48 tests
• 192 tests
• Bulk formats available for CROs and manufacturing partners

Ready to upgrade your T-cell assays?

The ClickTech EdU T-cell Proliferation Kit offers clear, direct insight into T-cell dynamics without the limitations of dilution dyes. Perfect assay for researchers in cancer immunology, infectious disease, autoimmunity, or cell therapy development.

LITERATURE

A sensitive and less cytotoxic assay for identification of proliferating T cells based on bioorthogonally-functionalized uridine analogue, F. C. Stempels et al., 2022, Journal of Immunological Methods, Vol. 502,  p. 113228.
https://doi.org/10.1016/j.jim.2022.113228