Apoptosis assay: In Situ Apoptosis Detection Kit
During apoptosis, DNA is fragmented following the apoptotic activation of intracellular endonucleases. The In Situ Apoptosis Detection Kit is designed to detect fragmented DNA histochemically by TUNEL (TdT-mediated dUTP Nick End Labeling). Via this method, fluorescein-labeled nucleotides are incorporated in situ onto the 3' ends of DNA fragments, allowing histologic localization and detection of individual apoptotic cells.
During apoptosis, DNA is fragmented following the apoptotic activation of intracellular endonucleases. The In Situ Apoptosis Detection Kit is designed to detect fragmented DNA histochemically by TUNEL (TdT-mediated dUTP Nick End Labeling). Via this method, fluorescein-labeled nucleotides are incorporated in situ onto the 3' ends of DNA fragments, allowing histologic localization and detection of individual apoptotic cells.
Apoptosis plays a significant role in various stages of cancer as well as cell development, growth, and proliferation. Physical (e.g., radiation, heat) and chemical (e.g., pharmacological) factors can also induce the apoptotic state. One of the features of apoptotic cells is the cleavage of nuclear DNA to approximately 185-bp fragments. This fragmented DNA can be detected histochemically by the TUNEL method.
Principle
TUNEL is an effective method for measuring DNA fragments resulting from the apoptotic activation of intracellular endonucleases. The TdT enzyme is used to label the 3'-OH ends of DNA fragments with fluorescein-dUTP. These fluorescence-labeled nucleotides are incorporated in situ onto DNA fragment ends, allowing histologic localization of individual cells. As a result, cells undergoing apoptosis can be easily detected by fluorescence microscopy. Since incorporated fluorescein-dUTP can also be detected using a peroxidase-labeled anti-fluorescein antibody, it is possible to detect apoptotic cells using light microscopy.
Overview
- Quick and easy detection of apoptotic cells
- Sensitive detection of single cells in the initial stages of apoptosis
- Specific staining of apoptotic but not necrotic cells
- Flexible enough to be used with tissue section or fixed-cell samples
- Individual components are available separately
- Control slide supplied for accuracy
- Non-radioactive
More Information
Applications
- Fragmented-DNA detection by the TUNEL method
- Detection of DNA strand breaks in apoptotic cells via fluorescence or light microscopy
Fragmented DNA detection by the TUNEL method
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Type: direct TUNEL labeling assay
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Used for: detection of DNA strand breaks in apoptotic cells by flow cytometry or fluorescence microscopy
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Used to assay: cells in suspension (permanent cell lines, normal and tumor cells ex vivo), adherent cells, cytospins, cell smears, frozen or paraffin-embedded tissue sections
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Technique: end-labeling of DNA with fluorescein-dUTP, followed by direct analysis of fluorescent cells
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Time required: 1–2 hr (Not including sample preparation, permeabilization, etc.)
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Detection method: fluorescence and light microscopy
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Number of Assays per Kit: 20
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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