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

Overview

  • Product nameIn situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit
    See all DNA fragments kits
  • Detection methodFluorescent
  • Tests
    60 x 1 assay
  • Sample type
    Tissue, Adherent cells, Suspension cells
  • Assay typeQuantitative
  • Assay time
    3h 00m
  • Product overview

    Abcam's In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit provides complete components including positive and negative control cells for conveniently detecting DNA fragmentation by fluorescence microscopy. The kit utilizes Br-dUTP (bromolated deoxyuridine triphosphate nucleotides) which is more readily incorporated into DNA strand breaks than other larger ligands (e.g., fluorescein, biotin or digoxigenin). The greater incorporation gives rise to brighter signal when the Br-dUTP sites are identified by a Red fluorescence labeled anti-BrdU monoclonal antibody. The assay is suitable for studying apoptosis with GFP transfected cells.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

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

  • Tested applicationsSuitable for: Fluorescence Microscopymore details

Properties

  • RelevanceInternucleosomal DNA fragmentation is a hallmark of apoptosis in mammalian cells.

Associated products

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
Fluorescence Microscopy Use at an assay dependent concentration.

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

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

This product has been referenced in:
  • Hopkins J  et al. Meiosis-specific cohesin component, Stag3 is essential for maintaining centromere chromatid cohesion, and required for DNA repair and synapsis between homologous chromosomes. PLoS Genet 10:e1004413 (2014). IHC-P ; Mouse . Read more (PubMed: 24992337) »

See 1 Publication for this product

Product Wall


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 ...

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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.
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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...

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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.
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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...

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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.
...

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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....

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Thank you for contacting us.

This kit should work for both cells in suspension and adherent cells.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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