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QIA21 FragEL™ DNA Fragmentation Detection Kit, Colorimetric - Klenow Enzyme

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

Synonyms: TUNEL Assay

QIA21
  
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      Overview

      Key Spec Table

      Detection Methods
      Colorimetric
      Description
      Overview

      This product has been discontinued.



      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 NumberQIA21
      Brand Family Calbiochem®
      SynonymsTUNEL 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
      ReferencesXiao, D. and Bullock, R. 1996. J. Neurosurgery 85, 655.
      Gu, Y., et al. 1994. Endocrinology 135, 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. Immunology 145, 1859.
      Wyllie, A. H. 1980. Nature 284, 555.
      Kerr, J. F. R., et al. 1972. Br. J. Cancer 26, 239.
      Product Information
      Detection methodColorimetric
      Form50 Tests
      FormatMicroscopy
      Kit containsCounterstain, 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.
      Quality LevelMQ100
      Applications
      Key Applications Frozen Sections
      Paraffin Sections
      Biological Information
      Assay time4 h
      Sample TypeFrozen or paraffin sections or fixed-cell preparations
      Species Reactivity
      • A Broad Range Of Species
      Product Usage Statements
      Intended useThe 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 ConditionsUpon 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
      Supplemental Information
      Kit containsCounterstain, 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.
      Global Trade Item Number
      Catalogue Number GTIN
      QIA21 0