Click-iT™ Plus EdU Alexa Fluor™ 488 Flow Cytometry Assay Kit is used to analyze DNA replication in proliferating cells , is a simpler and more reliable detection method than the traditional BrdU method. Analyze newly synthesized DNA using the 488 nm laser of a flow cytometer. Click-iT™ Plus formulations are compatible with standard fluorophores including R-PE, R-PE concatemers, and fluorescent proteins.

•Hybridable—compatible with R-PE (and concatemers) and fluorescent proteins
•Accurate—detection results are better than BrdU detection
•Fast—available in as little as 60 minutes Get results

View all flow cytometry assays Click- Selection Guide for iT™ EdU and Click-iT™ Plus EdU Assays.

Combinable
Compared to the original Click-iT™ EdU flow cytometry assay, the Click-iT Plus™ formulation offers better coupling capabilities. The Click-iT™ Plus EdU Assay can be used with R-PE and R-PE concatemers, as well as a variety of fluorescent proteins such as GFP and mCherry, while providing the accuracy and speed of the original Click-iT™ EdU Assay.

A better alternative to BrdU that provides better results
A more accurate proliferation assay that directly measures DNA synthesis. Initially, this was accomplished by incorporating radioactive nucleotides. Subsequently, the antibody-based bromodeoxyuridine (BrdU, nucleoside analog) method replaced this detection method. Click-iT™ Plus EdU flow cytometry assay is a new alternative to BrdU assay. EdU (5-ethynyl-2´-deoxyuridine) is a thymidine analog that is incorporated into DNA during active DNA synthesis. The detection is based on click chemistry, in which alkynes react with dye-labeled azides in a copper-catalyzed reaction to form stable covalent bonds. In this application, an alkyne is found in the acetylene group of EdU, while the azide is coupled to the Alexa Fluor™ dye. Standard flow cytometric analysis methods were used to determine the percentage of S-phase cells in the cell population.

Mild conditions for use with cell cycle dyes and antibodies
Small azide dye for efficient detection of incorporated EdU using mild conditions while having sufficient aldehyde-based Standard fixation and detergent permeabilization capabilities for use with Click-iT™ Plus detection reagents to obtain DNA. This is in contrast to BrdU detection, which requires the use of HCl, heat, or digestion of denatured DNA with Dnase to expose BrdU for detection with anti-BrdU antibodies. Treating samples with BrdU will change cell cycle distribution, and HCl can destroy antigen recognition sites. In contrast, the easy-to-use Click-iT™ Plus EdU assay is compatible with cell cycle dyes. The Click-iT™ Plus EdU Assay can also be used with antibodies to cell surface and intracellular markers as well as conjugates labeled with standard fluorophores including R-PE, R-PE concatemers, and fluorescent proteins (GFP and mCherry) .

Quick and easy protocol
Click-iT™ Plus EdU protocol is based on immunohistochemistry antibody labeling with aldehyde-based fixation and detergent permeabilization steps. However, EdU differs from others. Compatible with fixatives/permeabilizing agents including saponin and methanol. Analyze cell proliferation data in just five steps:

1. Treat cells with EdU.
2. Fix and permeabilize cells.
3. Use Click-iT™ Plus Assay Mix to detect S-phase cells for 30 minutes.
4. Wash once.

In some cases, within 60 minutes. Results are available but we recommend 90 minutes for all applications