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| Product Name | Fluor 594 EdU Cell Proliferation Assay Kit (Imaging & Flow Cytometry) |
| Catalog No. | RESK-0166 |
| Description | Cell proliferation analysis is a fundamental approach for assessing cell health, genotoxicity, and the efficacy of antitumor drugs. The BrdU assay is a commonly used method for detecting cell proliferation; the EdU assay is an upgraded and breakthrough improvement over the BrdU method. EdU (5-ethynyl-2'-deoxyuridine) is a pyrimidine nucleoside analog that is incorporated into DNA strands during DNA synthesis (S phase). EdU detection is based on click chemistry (copper-catalyzed azide-alkyne cycloaddition). |
| Intended Use | For the detection of cell proliferation and cell cycle analysis by fluorescence microscopy and flow cytometry. For research use only. |
| Principle / Technology | The EdU reagent in the kit contains an alkyne group, while the Fluor 594 Azide dye reagent contains an azide group. Incorporated EdU is labeled rapidly and efficiently through click chemistry (copper-catalyzed azide-alkyne cycloaddition), and only a small amount of the azide dye is needed to efficiently label the incorporated EdU. Standard aldehyde fixation and detergent permeabilization allow the detection reagents to enter the cells, whereas the BrdU method requires DNA denaturation (acid denaturation, heat denaturation, or DNase digestion) to expose BrdU for antibody binding. |
| Detection Method | Fluorescence microscopy and flow cytometry |
| Sample Type | Cultured proliferating cells |
| Performance Range / Specifications | Suitable for both cell proliferation detection and cell cycle analysis; compatible with fluorescence microscopy and flow cytometry. |
| Reaction Conditions / Protocol | 1. Label proliferating cells with EdU. 2. Fix cells with standard aldehyde fixation and permeabilize with detergent. 3. Perform the click reaction with the Fluor 594 Azide detection reagent. 4. Counterstain nuclei with Hoechst 33342 for cell cycle analysis if required. 5. Analyze by fluorescence microscopy or flow cytometry. |
| Components / Formulation | The kit contains all components required for EdU-based detection: Component A: 10 mM EdU Component B: Fluor 594 Azide Click reaction reagents, fixation/permeabilization reagents, and wash buffers Hoechst 33342 nuclear dye for cell cycle analysis |
| Storage Conditions | Store at -25 to -15 °C protected from light. After opening: Component A (10 mM EdU) at -25 to -15 °C; Component B (Fluor 594 Azide) at -25 to -15 °C protected from light; all other components at 2-8 °C. |
| Shelf Life | 1 year when stored at -25 to -15 °C protected from light. |
| Package Specifications | 100 T/10 T 500 T/50 T |
| Product Form | Kit |
| Concentration | Component A: 10 mM EdU |
| Expiration Date / Stability | Valid for 1 year at -25 to -15 °C protected from light. |
| Regulatory / Compliance | For research use only. |
| Application Notes / Precautions | Suitable for in vitro assays with proliferating cells capable of DNA synthesis; tissue samples are not applicable. Unlike BrdU immunodetection, which requires DNA denaturation before antibody binding, EdU carries an alkyne group that reacts directly with fluorescent dyes via click chemistry without denaturation, giving broader applicability. The assay cannot be performed in live cells: although EdU metabolically labels living cells, the azide detection reagent and buffer components are cell-impermeant, so the click reaction must be performed on fixed and permeabilized samples. High background is generally caused by insufficient washing; increase the number of BSA washes. Include a no-dye or no-click-reaction control processed under the same conditions to exclude autofluorescence, and perform a complete click reaction in an EdU-free system to verify signal specificity. If no signal or a very weak specific signal is observed: ensure reagents are colorless when used and do not use yellowed additives or buffers; ensure cells are fully fixed and permeabilized so the click reagents reach the nucleus; do not introduce metal chelators (e.g., EDTA, EGTA, citrate) into any buffer or reagent before the click reaction, as copper ions bind to them, and add extra washing steps for samples containing such substances; the click reaction requires copper ions in the correct valence state, Cu2+. Extending the click reaction beyond 30 minutes does not improve the signal; a fresh 30-minute incubation with fresh click reaction reagents improves labeling efficiency more effectively. |
For research use only, not for clinical use.
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