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.

     

    How other researchers have used HRP-DAB TUNEL Assay Kit ab206386

    This TUNEL assay kit has been used in publications in a variety of sample types, including:
    - Human: LX-2 cell cultures1 
    - Mouse (paraffin-embedded sections): abdominal aortic aneurysm lesion2, thyroid gland3, lung tissue4, heart5, liver6, ovary tissue7, skin8, liver9
    - Rat: paraffin embedded heart10, brain tissue11
    - Other: paraffin embedded human xenograft tumors in mice12, paraffin-embedded tissue from nude mice injected with human pancreatic cancer cell line13

    References: 1-Park SM et al 2017, 2-Li et al 2019, 3-Iglesias-Osma MC et al 2019, 4-Yuan X et al 2018, 5-Ravi V et al 2018, 6-Jadhav K et al 2018, 7-Gao Y et al 2017, 8-Shin JM et al 2017, 9-Kim MH et al 2017 and 2016, 10-Zhang J et al 2018, 11-Xiao B et al 2017, 12-Nielsen CF et al 2017, 13-Dauer et al 2019

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:

  • Li J  et al. IL (Interleukin)-33 Suppresses Abdominal Aortic Aneurysm by Enhancing Regulatory T-Cell Expansion and Activity. Arterioscler Thromb Vasc Biol 39:446-458 (2019). Read more (PubMed: 30651000) »
  • Dauer P  et al. ER stress sensor, glucose regulatory protein 78 (GRP78) regulates redox status in pancreatic cancer thereby maintaining "stemness". Cell Death Dis 10:132 (2019). Read more (PubMed: 30755605) »
See all 16 Publications for this product

Customer reviews and Q&As

Abreviews
Used on 4 µm paraffin sections human tonsil following instructions from the kit.

Abcam user community

Verified customer

Submitted Oct 12 2019

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