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manufacturer/tradename
Roche
storage condition
avoid repeated freeze/thaw cycles
General description
The DNA Fragmentation Imaging Kit is a simple and rapid TUNEL assay for detecting apoptosis induction in mammalian cells. DNA fragmentation, detected using terminal deoxynucleotidyl transferase and fluorescein-labeled dUTP, is measured by fluorescence microscopy or an automated imaging platform, such as the Cellavista System. The kit uses a dye to stain the nuclei of living and dead cells for quantifying total and apoptotic cell numbers, and calculating the percentage of apoptotic cells.
Application
Programmed cell death (apoptosis) is a key pathway in mammalian cells during tissue homeostasis. Cell death malfunction has been implicated in pathology. Quantification of caspase-triggered DNA fragmentation is one of the principal techniques used to detect apoptosis induction. The DNA Fragmentation Imaging Kit performs the TUNEL assay for accurate and fast quantitative fluorescence detection using terminal deoxynucleotidyl transferase and fluorescein-labeled dUTP. The kit is ideal for measuring apoptosis in medium to high throughput cellular workflows.
- Quickly determine apoptosis induction for cell-based workflows in life science research.
- Reliably perform screening assays in product development, e.g., cancer research, pre-clinical safety.
- Accurately measure malignant cell sensitivity in cancer research.
Features and Benefits
- Optimized for data evaluation with Roche′s Cellavista System. Obtain reproducible results using automated imaging with this system.
- Saves time and resources: Total assay time, including analysis, for a 96-well microplate is 85 minutes.
- Reduced risk of losing cells with fewer washing steps in an improved protocol.
Packaging
1 kit containing 3 components
Quality
The kit is function tested using a cellular model (HeLa cells treated with antimycin A).
Principle
Labeling of apoptotic cells
Genomic DNA cleavage during apoptosis produces double-stranded, low molecular weight DNA fragments (mono- and oligonucleosomes), as well as single strand breaks (nicks) in high molecular weight DNA. These DNA strand breaks are identified by labeling free 3′-OH termini with modified nucleotides in an enzymatic assay known as the TUNEL reaction. The TUNEL reaction preferentially labels DNA strand breaks generated during apoptosis. TUNEL assays discriminate apoptosis from necrosis, and from primary DNA strand breaks induced by cytostatic drugs or irradiation. The DNA Fragmentation Imaging Kit labels DNA strand breaks using terminal deoxynucleotidyl transferase (TdT). TdT catalyzes polymerization of labeled nucleotides to free 3′-OH DNA ends in a template-independent manner using the TUNEL reaction. Fluorescein incorporated in nucleotide polymers is detected and quantified using fluorescence microscopy.
Staining of nuclei
Hoechst 33342 is a cell-permeable stain that binds DNA. It is used to stain nuclei of living or dead cells.
Genomic DNA cleavage during apoptosis produces double-stranded, low molecular weight DNA fragments (mono- and oligonucleosomes), as well as single strand breaks (nicks) in high molecular weight DNA. These DNA strand breaks are identified by labeling free 3′-OH termini with modified nucleotides in an enzymatic assay known as the TUNEL reaction. The TUNEL reaction preferentially labels DNA strand breaks generated during apoptosis. TUNEL assays discriminate apoptosis from necrosis, and from primary DNA strand breaks induced by cytostatic drugs or irradiation. The DNA Fragmentation Imaging Kit labels DNA strand breaks using terminal deoxynucleotidyl transferase (TdT). TdT catalyzes polymerization of labeled nucleotides to free 3′-OH DNA ends in a template-independent manner using the TUNEL reaction. Fluorescein incorporated in nucleotide polymers is detected and quantified using fluorescence microscopy.
Staining of nuclei
Hoechst 33342 is a cell-permeable stain that binds DNA. It is used to stain nuclei of living or dead cells.
Preparation Note
Storage conditions (working solution): The Reaction Solution created by mixing the Enzyme Solution and Label Solution (Vial 2 and Vial 3), in Step 6 of the experiment protocol, cannot be stored. The Reaction Solution should be prepared just before use. Discard remaining unused Reaction Solution.
Storage and Stability
Store at 15 to 25 °C. (Protect from exposure to light. Store dry in dark.)
Other Notes
For life science research only. Not for use in diagnostic procedures.
Kit Components Only
Product No.
Description
- Nuclei Dye (blue cap), for fluorimetric determination of cell count based on labeling of cell nuclei
- Enzyme Solution (yellow cap), for labelling of free 3′-OH termini of DNA fragements with fluorescein dUTP
- Label Solution (clear cap), containing a nucleotid mix with fluorescein dUTP in reaction buffer
signalword
Danger
hcodes
Hazard Classifications
Aquatic Chronic 2 - Carc. 1B - Muta. 2 Inhalation
Storage Class
6.1D - Non-combustible, acute toxic Cat.3 / toxic hazardous materials or hazardous materials causing chronic effects
wgk_germany
WGK 3
flash_point_c
does not flash
Certificates of Analysis (COA)
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