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| Product Name | TUNEL Apoptosis Detection Kit (FITC) |
| Catalog No. | RESK-0169 |
| 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 (FITC) 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), FITC-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 labeling reaction of this kit has been optimized: an ideal ratio of FITC-12-dUTP to unlabeled dNTP is used for nucleotide incorporation at 3'-OH termini, so that a longer 'labeled tail' can form at the end of each fragmented DNA strand. This labeled tail reduces steric hindrance between the label groups on adjacent incorporated dNTPs, increases the number of fluorophores on each DNA fragment, and minimizes aggregation and quenching that may occur when fluorophores are too close together, thereby improving detection sensitivity and reducing non-specific reactions. |
| 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 FITC-12-dUTP at the 3'-hydroxyl (3'-OH) termini exposed at genomic DNA breaks generated during apoptosis. |
| Detection Method | Fluorescence microscopy or flow cytometry (FITC fluorescence). |
| 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. Optimized labeling chemistry improves detection sensitivity and reduces non-specific reactions. |
| Reaction Conditions / Protocol | Fix samples with freshly prepared 4% neutral paraformaldehyde at 4 °C for approximately 25 min. Permeabilization is required after fixation so that the TdT enzyme can enter the cells. Do not prolong fixation excessively, as over-crosslinking reduces labeling efficiency; avoid ethanol, methanol, and acidic fixatives, which lead to low labeling efficiency or false positives. 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 flow cytometric TUNEL detection, PI staining is not mandatory and is usually omitted. |
| Components / Formulation | FITC-12-dUTP Labeling Mix Terminal Deoxynucleotidyl Transferase (TdT) |
| Storage Conditions | Store the kit at -20 °C. The FITC-12-dUTP Labeling Mix should be stored at -20 °C protected from light. |
| Shelf Life | 2 years |
| Package Specifications | 20 T 50 T 100 T |
| Product Form | Kit |
| Shipping Conditions | Shipped on wet ice. |
| Expiration Date / Stability | Stable for 2 years when stored as recommended. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Compatible with DAPI co-staining. In flow cytometric TUNEL detection, PI counterstaining is optional. |
| Application Notes / Precautions | If the TUNEL signal becomes diffuse and loses nuclear localization after co-staining, excessive washing may be the cause; reduce the number of washing steps and wash gently. TUNEL staining can be performed together with DAPI staining. Annexin V detects early apoptosis, JC-1 detects mid-stage apoptosis, and TUNEL detects late-stage apoptosis. Applicable to plant cells or bacteria (prokaryotes), but protoplasts must first be prepared because these cells have cell walls; the staining solution only needs to cover the cells, and staining time varies among cell types. |
For research use only, not for clinical use.
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