Product nameTUNEL Assay Kit - HRP-DAB
See all DNA fragmentation kits
Sample typeTissue, Adherent cells
Assay typeCell-based (qualitative)
Assay time5h 0m
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
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.
Storage instructionsStore 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
RelevanceInternucleosomal DNA fragmentation is a hallmark of apoptosis in mammalian cells.
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)
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) »