TUNEL Assay Kit - In situ BrdU-Red DNA Fragmentation (ab66110)

Overview

  • Product name
    TUNEL Assay Kit - In situ BrdU-Red DNA Fragmentation
    See all DNA fragments kits
  • Detection method
    Fluorescent
  • Sample type
    Tissue, Adherent cells, Suspension cells
  • Assay type
    Quantitative
  • Assay time
    3h 00m
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Mammal
  • Product overview

    TUNEL Assay Kit - In situ BrdU-Red DNA Fragmentation (ab66110) is a convenient and sensitive method to detect DNA fragmentation by flow cytometry and fluorescence microscopy in live cells. The kit uses Br-dUTP (bromolated deoxyuridine triphosphate nucleotide), which can be more readily incorporated into DNA strand breaks that other dUTP labels such as fluorescein, biotin or dioxigenin. The greater incorporation rate produces a brighter signal when the Br-dUTP sites are detected with an anti-BrdU monoclonal antibody directly labeled with a red fluorochrome.


    The kit includes cells that can be used as positive or negative control. This assay is especially suitable for studying DNA fragmentation in GFP-transfected cells.



    Visit our FAQs page for tips and troubleshooting.

  • Notes

    This kit is BrdU-Red labeled (Ex/Em = 488/576 nm). If want to use FITC (Ex/Em = 495/519 nm) as a label, we recommend our In situ Direct DNA Fragmentation (TUNEL) Assay Kit (ab66108).

  • Tested applications
    Suitable for: FM, Flow Cyt, IHC-Pmore details

Properties

Applications

Our Abpromise guarantee covers the use of ab66110 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
FM Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. PubMed: 24992337

Images

  • Immunohistochemical analysis of paraffin embedded 5 micron thick testis tissues of 8 week old Stag3+/− and Stag3−/− (Stromal antigen) mice . Apoptotic cells were detected using In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (ab66110). DAPI was used as a counterstain.

  • Detection of DNA fragmention (TUNEL staining) using the negative and positive control cells (HL-60 untreated and treated with camptothecin). Cells were stained following the assay protocol. The fluorescence signal was detected and analyzed using BD FACScan System (Becton Dickinson).

  • TUNEL staining in whole mount Hydractinia echinata using In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (ab66110).

    Animals were fixed in 4% PFA in PBS for 1 hour and processed as per the protocol without proteinaseK treatment. In place of proteinaseK animals were permeabilised in 3% Triton in PBS for 15 minutes. Animals were counter-stained with DAPI.

Protocols

References

This product has been referenced in:
  • Jin WN  et al. Depletion of microglia exacerbates postischemic inflammation and brain injury. J Cereb Blood Flow Metab 37:2224-2236 (2017). IHC-P ; Mouse . Read more (PubMed: 28273719) »
  • Yang S  et al. Probucol ameliorates renal injury in diabetic nephropathy by inhibiting the expression of the redox enzyme p66Shc. Redox Biol 13:482-497 (2017). Read more (PubMed: 28728079) »

See all 4 Publications for this product

Customer reviews and Q&As

Abreviews
I purchased this kit with the purpose of staining cultured cells. It totally failed in that purpose and I would recommend the xxxx kit if your aim is to investigate apoptosis in cultured cells.
The protocol provided by Abcam does not mention any suggestion for cultured cells.
Additionally, the protocol is not well written. This includes a mistake on the manual with a recommended amount to pipette. You will easily find the mistake but this should not happen. I sent an email to Abcam and I hope they correct the mistake.

On the other hand, the kit works pretty well for tissues embedded in Paraffin. Attached is an example of mouse mammary gland. Magnification = 20X
Username

Abcam user community

Verified customer

Submitted Sep 08 2017


We removed flow cytometry as an application for ab66110 In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Ki several years ago. We had received feedback that there was a lot of background staining and it also failed a retest in our laboratories. I ...

Read More

Dual Caspase-3/TUNEL Fluorescence Staining

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Frozen sections of snap frozen sekeletal muscle tissue with isopentane were used in the application. Sections were fixed in cold acetone for 10 minutes and then treated with 5% normal goat serum as a blocking reagent for 30 minutes.

*PBS was used for rinsing/washing solution in between steps.

No antigen retrieval used.

PART A: Caspase-3
i. Primary antibody ab52293 anti-Caspase-3 (1/200, overnight incubation at 4C)
ii. Secondary antibody ab7086 Goat anti-Rabbit IgG H&L FITC (1/200, 1 hour incubation at RT)

PART B: TUNEL
i. As per kit protocol from Part 2 (Detection by Fluorescence Microscopy), steps (a) to (l)
ii. After step (l), sections were dipped in DAPI diluted in PBS (1/10000) for 1 minute prior to final washing steps

Sections were coverslipped using fluoromount mountant and kept refridgerated at 4C in the dark until ready for visualisation.
Username

Abcam user community

Verified customer

Submitted Apr 07 2016

We have not done any additional antibody staining with the ab66110 In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit .Whether it can be done will depend on the criteria required by the additional antibody but theoretically staining should work afte...

Read More
Abreviews
Animals were fixed in 4% PFA in PBS from 1 hour and processed as per the protocol without proteinaseK treatment. In place of proteinaseK animals were permeabilised in 3%Triton in PBS for 15 minutes. Animals were counter-stained with DAPI.
Username

Mr. James Gahan

Verified customer

Submitted Oct 06 2014

This kit is specifically designed for and has been used for co detection of GFP transfected cells and the Red fluorescence labeled anti-BrdU monoclonal antibody. There is therefore no need to use this other antibody conjugated to AlexaFluor647.
<...

Read More



The 7AAD stain has different excitation and emission maxima than the Red fluorescence from the antibody ( emission max is 576 versus 655nm). This difference is enough to distinguish spectrally the signal from the antibody and 7AAD. The main r...

Read More

J’ai contacté le laboratoire qui m’a confirmé que pour la microscopie, nous vous recommandons de prendre les cellules et d’ajouter du PBS avant d'ajouter le 7-AAD et d’observer les cellules sous un microscope.
...

Read More

J’ai contacté le laboratoire qui m’a confirmé que les cellules pour les contrôles positifs et négatifs sont déjà fixés et sont stockés dans 70% éthanol.



The lab have confirmed that we have never measured DFI with this kit. However, my online search seems to indicate that it is possible to do so. Here is the formula and the link for its calculation, which needs to be modified according to this staining....

Read More

1-10 of 13 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up