Thermo Fisher Scientific Click-iT Plus EdU Alexa Fluor 594 Flow Cytometry Assay Kit

Invitrogen™ Click-iT™ Plus EdU Alexa Fluor™ 594 Flow Cytometry Assay Kit

The Click-iT Plus EdU Alexa Fluor 594 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 532- or 561-nm laser of a flow cytometer. The Click-iT Plus formulation is compatible with standard fluorophores including R-PE, R-PE tandems, and fluorescent proteins.

주요 특징

• Multiplexable: Compatible with R-PE (and tandems) and fluorescent proteins such as GFP and mCherry
• 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

Click-iT Plus Alexa Fluor 594 EdU assays can be used with R-PE and R-PE tandems, as well as fluorescent proteins such as GFP and mCherry. The Alexa Fluor 594 labeling reagent is excited at 532 or 561 nm and emits at 615 nm.

Advanced Method Superior to BrdU

EdU (5-ethynyl-2´-deoxyuridine) is a thymidine analog incorporated into DNA during active synthesis. Detection uses click chemistry—a copper-catalyzed covalent reaction between an azide and an alkyne. The alkyne is in EdU’s ethynyl group, and the azide is coupled to the Alexa Fluor dye. Standard flow cytometry methods determine the percentage of S-phase cells.

Mild Conditions Compatible with Dyes and Antibodies

The small dye azide allows efficient detection of incorporated EdU under mild conditions. Standard aldehyde fixation and detergent permeabilization are sufficient for detection, unlike BrdU assays that require DNA denaturation (HCl, heat, or DNase). The Click-iT Plus EdU assay maintains cell cycle signal integrity and antigen recognition, and is compatible with cell cycle dyes and antibody multiplexing.

Quick and Simple Protocol

The Click-iT Plus EdU protocol follows standard fixation and permeabilization steps similar to immunohistochemical labeling. EdU is also compatible with saponin and methanol treatments.

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 (recommended 90 minutes for all applications).

제품 사양