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| Product Name | TUNEL Apoptosis Detection Kit (Fluor 488) |
| Catalog No. | RESK-0170 |
| Description | When cells undergo apoptosis, endogenous DNA endonucleases are activated and cleave genomic DNA between nucleosomes; DNA extracted from apoptotic cells shows a characteristic 180-200 bp DNA ladder upon electrophoresis. The TUNEL Apoptosis Detection Kit (Fluor 488) is used to detect nuclear DNA fragmentation during late-stage apoptosis in tissue and cultured cells. Its principle is that, under the action of Terminal Deoxynucleotidyl Transferase (TdT), Fluor 488-12-dUTP is incorporated at the 3'-hydroxyl (3'-OH) termini exposed at genomic DNA breaks, allowing detection by fluorescence microscopy or flow cytometry. The Fluor 488 dye offers higher stability and a stronger signal, making the label brighter and more resistant to quenching. |
| Intended Use | Detection of nuclear DNA fragmentation in late-stage apoptosis of tissue and cultured cells. |
| Principle / Technology | TUNEL (TdT-mediated dUTP nick end labeling). Terminal Deoxynucleotidyl Transferase (TdT) catalyzes the incorporation of Fluor 488-12-dUTP at the 3'-hydroxyl (3'-OH) termini exposed at genomic DNA breaks generated during apoptosis. |
| Detection Method | Fluorescence microscopy, confocal microscopy, flow cytometry, or fluorescence microplate reader. |
| Sample Type | Frozen or paraffin-embedded tissue sections; cultured adherent or suspension cells. |
| Performance Range / Specifications | Broad applicability: suitable for detecting apoptosis in frozen or paraffin-embedded tissue sections and in cultured adherent or suspension cells. The Fluor 488 dye provides higher stability, stronger signal, brighter labeling, and greater resistance to quenching. |
| Reaction Conditions / Protocol | Fix samples with freshly prepared 4% neutral paraformaldehyde; ethanol or methanol fixation results in low labeling efficiency, and acidic fixatives may cause false positives. Permeabilization is required because the TdT enzyme must enter the cells to reach the DNA breaks. Adherent cells do not need to be detached before staining: wash 2-3 times with PBS, fix directly with paraformaldehyde, permeabilize, then stain and observe. For coverslip (slide) observation, add enough staining working solution to fully cover the cells; for flow cytometry, refer to the manual for the recommended working solution volume. Observe samples immediately after staining; the entire observation should be completed within 30 min. |
| Components / Formulation | Fluor 488-12-dUTP Labeling Mix Terminal Deoxynucleotidyl Transferase (TdT) |
| Storage Conditions | Store the kit at -20 °C. The Fluor 488-12-dUTP Labeling Mix should be stored at -20 °C protected from light. |
| Shelf Life | 1 year |
| Package Specifications | 20 T 50 T 100 T |
| Product Form | Kit |
| Shipping Conditions | Shipped on wet ice. |
| Expiration Date / Stability | Stable for approximately 1 year when stored under the recommended conditions. |
| Regulatory / Compliance | For research use only. |
| Compatibility | TUNEL fluorescent signals can be detected by fluorescence microscopy, confocal microscopy, flow cytometry, and microplate readers. Applicable to plant cells or bacteria (prokaryotes) after protoplast preparation. |
| Recommended Buffer System | If a dilution buffer is supplied with the kit, use it to prepare the staining working solution; if not, an appropriate buffer such as HBSS, serum-free medium, or PBS can be used. |
| Application Notes / Precautions | Fluorophores undergo severe quenching upon prolonged exposure to ordinary light; perform handling and incubation protected from light. Results can be read on a multifunctional microplate reader, although fluorescence microscopy, confocal microscopy, or flow cytometry is recommended when available. If fluorescence is weak, appropriately extend the incubation time or increase the working solution concentration, and use cells in good condition. If fluorescence quenches rapidly after excitation, the reagent concentration may be too high; reduce the concentration, use a low-energy laser to locate the field of view before switching to high-energy excitation, or use an anti-quenching (anti-fade) reagent. Unlike Annexin V and JC-1 assays, which cannot be performed on fixed cells, TUNEL samples require fixation and permeabilization. |
For research use only, not for clinical use.
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