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Description
When cells undergo apoptosis, they activate endonuclease enzymes that cut genomic DNA between nucleosomes. DNA ladder of 180-200 bp can be detected by electrophoresis after extraction of DNA during apoptosis.
TUNEL (TdT mediated dUTP Nick End Labeling) Cell apoptosis Assay Kit (FITC) can be used to detect the nuclear DNA breakage in tissue cells during late apoptotic stage. The principle is that FITC-12-dUTP is inserted into the 3'-hydroxyl (3'-OH) terminal of genomic DNA exposed during DNA break under the action of Terminal Deoxynucleotidyl Transferase (TdT). This can be detected by fluorescence microscopy or flow cytometry.
This kit optimizes the labeling reaction by using FITC-12-dUTP and unlabeled dNTP at the optimal ratio for nucleotide incorporation at the 3' -OH terminal, resulting in a longer "labeling tail" at the end of the same broken DNA fragment. The “labeling tail” reduces the steric hindrance of labeled groups on adjacent dNTP, increases the number of fluorescent-groups on each broken fragment, and reduces the aggregation and quenching that may be caused by adjacent fluorescent-groups, thus improving detection sensitivity and reducing non-specific reactions.
This kit has a wide range of applications. It can be used to detect apoptosis in frozen or paraffin sections, as well as in culture adherent cells or suspension cells situation.
Features
- The reagents in this kit are provided in liquid form for easy use.
- The kit can be used for a variety of purposes: flow cytometry, fluorescence microscopy for detection.
- Under the fluorescence microscope, the color distinction is obvious.
Applications
- This kit can be used for flow cytometry and fluorescence microscopy.
- This kit can detect the early apoptosis of cells.
- This kit can distinguish between late apoptotic and necrotic cells.
Specifications
| Application | The verified application of the kit was to detect apoptotic cells by flow cytometry or immunofluorescence cytology. |
| Packing form | Bottle |
Components
| Component | Name | 40306ES20 (20T) | 40306ES50 (50T) | 40306ES60 (100T) |
| 40306-A | 5×Equilibration Buffer | 750 μL | 1.25 mL×2 | 1.25 mL×3 |
| 40306-B | FITC-12-dUTP Labeling Mix | 100 μL | 250 μL | 250 μL×2 |
| 40306-C | Recombinant TdT Enzyme | 20 μL | 50 μL | 50 μL×2 |
| 40306-D | Proteinase K (2 mg/mL) | 40 μL | 100 μL | 100 μL×2 |
| 40306-E | DNase I (1 U/ μL) | 5 μL | 12.5 μL | 25 μL |
| 40306-F | 10 × DNase I Buffer | 100 μL | 250 μL | 500 μL |
Shipping and Storage
The product is shipped with dry ice and can be stored at -15℃ ~ -25℃ for 1 year.
Documents:
Citations & References:
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During early apoptosis, phosphatidylserine (PS) translocates from the inner to the outer leaflet of the plasma membrane before membrane integrity is lost. Annexin V binds PS with high affinity in a calcium-dependent manner, enabling early-apoptosis detection. Co-staining with propidium iodide (PI) discriminates: Annexin V⁺/PI⁻ = early apoptosis; Annexin V⁺/PI⁺ = late apoptosis/necrosis; Annexin V⁻/PI⁻ = viable cells.
TUNEL (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) detects DNA strand breaks — a late-stage hallmark of apoptosis — by enzymatically labeling 3′-OH termini with fluorescent dUTP. TUNEL is preferred for detecting apoptosis in fixed cells, tissue sections, or when intracellular markers are needed (e.g., co-staining with nuclear markers). Annexin V is suitable for live, intact cells analyzed by flow cytometry or fluorescence microscopy.
Channel requirements depend on the kit format: FITC variants require a 488 nm laser with 530/30 nm BP filter; YSFluor 488 variants are similar; YSFluor 647 requires a 633/647 nm (red) laser with 660/20 nm or 670 LP filter. Confirm your cytometer's laser and filter configuration matches the kit's excitation/emission profile. PI (propidium iodide) is typically detected in the PE or PerCP channel (488 nm excitation, 610–620 nm emission).
Avoid mechanical or chemical stress during harvest — use gentle enzymatic dissociation (Accutase preferred over trypsin) and keep cells at 4°C during staining. Wash with cold Annexin V Binding Buffer (calcium-containing) immediately before staining. Do not fix cells before Annexin V staining — fixation disrupts membrane phospholipid asymmetry. Process and analyze samples within 1 hour of staining to prevent signal drift.
Yeasen Biotechnology offers flexible customization options for many of its assay kits and detection reagents, including custom lot sizes, bulk ordering, and application-specific formulation adjustments. Volume pricing, custom packaging, and kit bundling may be available depending on the product and intended workflow. A Certificate of Analysis (CoA) and lot-specific QC data are provided with every order. For inquiries regarding large-volume orders, custom configurations, or integration into automated workflows, please contact the BioHippo team for a tailored quotation.
