Non-isotopic kit for the detection of apoptosis at the individual cell level. Apoptotic morphology is easily detected. This kit allows the recognition of apoptotic nuclei in paraffin-embedded tissue sections, tissue cryosections, or in cell preparations fixed on slide by Fragment End Labeling (FragEL™) of DNA. Klenow binds to exposed ends of DNA fragments generated in response to apoptotic signals and catalyzes the template-dependent addition of biotin-labeled and unlabeled deoxynucleotides. Biotinylated nucleotides are detected using streptavidin-horseradish peroxidase (HRP) conjugate. Diaminobenzidine reacts with the labeled sample to generate an insoluble colored product at the site of DNA fragmentation. Counterstaining with methyl green aids in the morphological evaluation and characterization of normal and apoptotic cells.
Catalogue Number
QIA21
Brand Family
Calbiochem®
Synonyms
TUNEL Assay
Materials Required but Not Delivered
• xylene (for paraffin sections only) • ethanol, 100, 90, 80, 70% (for paraffin sections only) • methanol • 30% hydrogen peroxide • tris-buffered saline (1X TBS, 20 mM Tris pH 7.6, 140 mM NaCl) • 10 mM Tris pH 8 • 1 mM MgSO4 in 1X TBS (optional, for use in generating positive control) • DNaseI (optional, for use in generating positive control) • distilled water • coplin jars, glass or plastic with slide holders • wash bottle or beaker for rinsing slides • humidified chamber • glass coverslips • mounting media (such as Permount®) • microscope • 2-20 µl, 20-200 µl, and 200-1000 µl precision pipetters with disposable tips • microcentrifuge tubes • parafilm® • absorbent wipes • ice
References
References
Xiao, D. and Bullock, R. 1996. J. Neurosurgery85, 655. Gu, Y., et al. 1994. Endocrinology135, 1272. Gavrieli, Y., et al. 1992. J. Cell Biol.119, 493. Fawthrop, D. J., et al. 1991. Arch. Toxicol.65, 437. Martin, S.J., et al. 1990. J. Immunology145, 1859. Wyllie, A. H. 1980. Nature284, 555. Kerr, J. F. R., et al. 1972. Br. J. Cancer26, 239.
Product Information
Detection method
Colorimetric
Form
50 Tests
Format
Microscopy
Kit contains
Counterstain, Proteinase K, Equilibration and Labeling Buffers, Klenow Enzyme (Cat. No. PF059), Stop buffer, Blocking Buffer, 50X Conjugate, DAB, Positive and Negative Control Slides, H₂O₂/Urea Tablets, Methyl Green Counterstain, and a user protocol.
Applications
Key Applications
Frozen Sections Paraffin Sections
Biological Information
Assay time
4 h
Sample Type
Frozen or paraffin sections or fixed-cell preparations
Species Reactivity
A Broad Range Of Species
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Intended use
The Calbiochem® Klenow-FragEL™ DNA Fragmentation Detection Kit is a non-isotopic system for the labeling of DNA breaks in apoptotic nuclei in paraffin-embedded tissue sections, tissue cryosections, and in cell preparations fixed on slides.
Storage and Shipping Information
Ship Code
Dry Ice Only
Toxicity
Multiple Toxicity Values, refer to MSDS
Hazardous Materials Attention:
Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.
Storage
-20°C
Storage Conditions
Upon arrival store the entire contents of the kit at -20°C in a non-frostfree freezer. Following initial thaw store the Stop Buffer and Methyl Green Counterstain at room temperature and the Blocking Buffer at 4°C. Refreezing of these components, however, does not appear to affect their performance.
Avoid freeze/thaw
Avoid freeze/thaw
Do not freeze
Ok to freeze
Packaging Information
Transport Information
Supplemental Information
Kit contains
Counterstain, Proteinase K, Equilibration and Labeling Buffers, Klenow Enzyme (Cat. No. PF059), Stop buffer, Blocking Buffer, 50X Conjugate, DAB, Positive and Negative Control Slides, H₂O₂/Urea Tablets, Methyl Green Counterstain, and a user protocol.
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