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

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

The Click-iT™ Plus EdU Pacific Blue™ Flow Cytometry Assay Kit provides a simplified and robust method for analyzing DNA replication in proliferating cells, offering superior performance compared to traditional BrdU assays. Newly synthesized DNA is analyzed using the 405 nm laser of the flow cytometer. The Click-iT™ Plus formulation is compatible with standard fluorophores, including R-PE and R-PE tandems, as well as fluorescent proteins.

주요 특징

• Multiplexable — Compatible with R-PE (and tandems) and fluorescent proteins
• Accurate — Provides superior results compared to BrdU assays
• Fast — Results in as little as 60 minutes

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

Multiplexable

The Click-iT™ Plus formulation provides increased multiplexibility compared to the original Click-iT™ EdU Flow Cytometry assays. It can be used in conjunction with R-PE, R-PE tandems, and fluorescent proteins such as GFP and mCherry without loss of accuracy or speed.

Advanced Method Superior to BrdU

Direct measurement of DNA synthesis is the most accurate method for proliferation analysis. The Click-iT™ Plus EdU assay replaces BrdU-based methods with a click chemistry approach, using EdU (5-ethynyl-2´-deoxyuridine) as a thymidine analog incorporated during DNA synthesis. Detection occurs through a copper-catalyzed reaction between an azide (coupled to the Alexa Fluor™ dye) and the alkyne group in EdU. Standard flow cytometry methods are then used to determine S-phase cell populations.

Mild Conditions for Compatibility

The small dye azide enables efficient EdU detection under mild conditions using standard aldehyde-based fixation and detergent permeabilization. Unlike BrdU assays, this method avoids harsh DNA denaturation (HCl, heat, or DNase), preventing signal alteration and preserving antigen recognition. The Click-iT™ Plus EdU assay is compatible with cell cycle dyes, antibodies, and standard fluorophores including R-PE, R-PE tandems, GFP, and mCherry.

Quick and Simple Protocol

The Click-iT™ Plus EdU protocol is easy to follow and compatible with multiple fixation and permeabilization agents (aldehyde, saponin, methanol).

5단계 절차

1. Treat cells with EdU
2. Fix and permeabilize cells
3. Detect S-phase cells with Click-iT™ Plus detection cocktail (30 min)
4. Wash once
5. Analyze

Results can be obtained in as little as 60 minutes, though 90 minutes is recommended for optimal performance.

사양 (Specifications)