Thermo Fisher Scientific Click-iT EdU Pacific Blue Flow Cytometry Assay Kit

Invitrogen™ Click-iT™ EdU Pacific Blue™ Flow Cytometry Assay Kit

The Click-iT™ EdU Pacific Blue™ Flow Cytometry Assay Kit provides a simplified and robust assay for analyzing DNA replication in proliferating cells compared to traditional BrdU methods. Newly synthesized DNA is analyzed using the 405 nm laser of a flow cytometer.

• Accurate — superior results compared to BrdU assays
• Fast — results in as little as 90 minutes
• Economical — more assays per kit

View selection guide for all Click-iT™ EdU and Click-iT™ Plus EdU assays for flow cytometry

An Advanced Method Giving You Results Superior to BrdU

The most accurate method of proliferation analysis is the direct measurement of DNA synthesis. Traditionally, this was performed using radioactive nucleosides such as 3H-thymidine, later replaced by antibody-based detection of bromodeoxyuridine (BrdU).
The Click-iT™ EdU Flow Cytometry Assay Kits are novel alternatives using EdU (5-ethynyl-2´-deoxyuridine), a thymidine analog incorporated into DNA during active synthesis. Detection is based on click chemistry — a copper-catalyzed covalent reaction between an azide and an alkyne.
In this assay, the alkyne is part of EdU, while the azide is coupled to Pacific Blue™, Alexa Fluor™ 488, or Alexa Fluor™ 647 dyes. Standard flow cytometry methods are then used to determine the percentage of S-phase cells.

Mild Conditions Allow Use with Cell Cycle Dyes and Antibodies

The small size of the dye azide allows efficient detection of incorporated EdU under mild conditions. Standard aldehyde-based fixation and detergent permeabilization are sufficient for reagent access to DNA.
Unlike BrdU assays, which require DNA denaturation (HCl, heat, or DNase digestion), the EdU method preserves cell cycle signal integrity and antigen recognition sites.
This compatibility allows multiplexing with cell cycle dyes and antibodies against surface or intracellular markers.

Quick and Simple Protocol

The Click-iT™ EdU protocol is based on standard fixation and permeabilization steps used in immunohistochemistry and is compatible with saponin and methanol.
In just five steps, you can analyze your cell proliferation data:

1. Treat cells with EdU
2. Fix and permeabilize cells
3. Detect S-phase cells with Click-iT™ detection cocktail for 30 minutes
4. Wash once
5. Analyze by flow cytometry

Get Accurate Results Economically

With more assays per kit, the Click-iT™ EdU Pacific Blue™ Flow Cytometry Assay Kit is more cost-effective than traditional BrdU assays, making it ideal for large-scale experiments.

For Research Use Only. Not for use in diagnostic procedures.

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