Overview

  • Product name
    TUNEL Assay Kit - HRP-DAB
    See all DNA fragmentation kits
  • Detection method
    Colorimetric
  • Sample type
    Tissue, Adherent cells
  • Assay type
    Cell-based (qualitative)
  • Assay time
    5h 0m
  • Product overview

    TUNEL Assay Kit - HRP-DAB (ab206386) (previously called In situ Apoptosis Detection Kit (DAB)) allows the recognition of apoptotic nuclei in paraffin-embedded tissue sections, frozen tissue sections, or in preparations of single cell suspensions fixed on slides.


    In the TUNEL assay protocol:
    - terminal deoxynucleotidyl Transferase (TdT) binds to exposed 3’-OH ends of DNA fragments generated in response to apoptotic signals and catalyzes the addition of biotin-labeled deoxynucleotides
    - biotinylated nucleotides are bound with a streptavidin-horseradish peroxidase (HRP) conjugate
    - diaminobenzidine (DAB) reacts with the HRP labeled sample to generate an insoluble colored (brown) substrate at the site of DNA fragmentation 
    - counterstaining with methyl green aids in the evaluation of normal and apoptotic cells

  • Notes

    This kit is designed for chromogenic TUNEL staining with HRP and DAB.

    To use FITC (Ex/Em = 495/519 nm) as a label, we recommend TUNEL Assay Kit - FITC (ab66108).

    To use BrdU-Red (Ex/Em = 488/576nm) as a label, we recommend TUNEL Assay Kit - BrdU-Red (ab66110).

    Find out more about the TUNEL method in the TUNEL staining / TUNEL assay guide.

     

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 30 slides 60 slides
    25X Conjugate 1 x 150µl 1 x 300µl
    Blocking Buffer 1 x 12ml 1 x 24ml
    DAB Solution 1 (DAB Concentrate) 1 x 150µl 1 x 300µl
    DAB Solution 2 (Substrate Reaction Buffer) 1 x 4ml 1 x 8ml
    Methyl Green Counterstain 1 x 3.5ml 2 x 3.5ml
    Proteinase K 1 x 50µl 1 x 100µl
    Stop Buffer 1 x 4ml 1 x 8ml
    TdT Enzyme 1 x 40µl 1 x 70µl
    TdT Equilibration Buffer 1 x 4ml 1 x 8ml
    TdT Labeling Reaction Mix 1 x 1.3ml 2 x 1.3ml
  • Research areas
  • Relevance
    Internucleosomal DNA fragmentation is a hallmark of apoptosis in mammalian cells.

Images

  • Gao Y et al used In situ Apoptosis Detection Kit / TUNEL assay ab206386 to analyze tissue sections from mouse ovaries.

    a. Section treated with DNase I as positive control

    b. Negative control without TdT enzyme

    c and f. representative experimental images.

    Nuclei stained with the TUNEL assay are brown. Sections were counter-stained with Methyl Green.

  • Using paraffin fixed human tonsil tissue,10 μm sections (1000X). A] Section processed and counter-stained with methyl green according to the manual. B] Counter-stain step was eliminated to more clearly illustrate the level of positive staining in the germinal centres of tonsil tissue. C] Section treated with DNase I in order to generate a positive control slide. Note all nuclei stain positive. The use of DNase I generates free 3’-OH groups on cellular DNA, these free 3’-OH groups are then labelled with biotin-nucleotide by the TdT in the kit. D] Negative control, the TdT enzyme step was eliminated thereby generating a negative slide.

  • Using paraffin fixed human tonsil tissue, 10 μm sections (1000X)

Protocols

References

This product has been referenced in:
  • Jadhav K  et al. Reversal of metabolic disorders by pharmacological activation of bile acid receptors TGR5 and FXR. Mol Metab 9:131-140 (2018). Read more (PubMed: 29361497) »
  • Yuan X  et al. Allergy immunotherapy restores airway epithelial barrier dysfunction through suppressing IL-25 -induced endoplasmic reticulum stress in asthma. Sci Rep 8:7950 (2018). Read more (PubMed: 29784924) »
See all 13 Publications for this product

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