Overview

  • Product name
    TUNEL Assay Kit - BrdU-Red
    See all DNA fragmentation kits
  • Detection method
    Fluorescent
  • Sample type
    Tissue, Adherent cells, Suspension cells
  • Assay type
    Quantitative
  • Assay time
    3h 00m
  • Species reactivity
    Reacts with: Other species, Mammals
  • Product overview

    TUNEL Assay Kit - BrdU-Red ab66110 (previously called TUNEL Assay Kit - In situ BrdU-Red DNA Fragmentation) uses a convenient and sensitive method to detect DNA fragmentation by flow cytometry and fluorescence microscopy in live cells.


    This TUNEL assay protocol uses Br-dUTP (bromolated deoxyuridine triphosphate nucleotide), which can be more readily incorporated into DNA strand breaks by the TdT enzyme than 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.


    BrdU-Red can be analyzed at Ex/Em 488/576 nm, with an optional 7-AAD counterstain at Ex/Em 488/655nm.


    The TUNEL assay kit includes both negative and postive control cells. It is designed to be suitable for studying DNA fragmentation in GFP-transfected cells.


    Tunel assay protocol summary:
    - fix cells / tissues with formaldehyde, or deparaffinize and rehydrate if paraffin sections, and wash
    - incubate cells in 70% ethanol for 30 min at 4ºC, or if tissues incubate with proteinase K solution for 5 min at room temp and refix with formaldehyde
    - wash
    - incubate in DNA labeling solution for 60 min at 37ºC
    - wash
    - incubate in antibody solution for 30 min at room temp
    - add 7-AAD / RNase A solution and incubate for 30 min at room temp
    - analyze with flow cytometry or fluorescent microscopy

  • Notes

    This kit is BrdU-Red labeled (Ex/Em = 488/576 nm).

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

    For chromogenic TUNEL staining, we recommend TUNEL Assay Kit - HRP-DAB ab206386.

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

  • Platform
    Flow cytometer, Fluorescence microscope

Properties

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:
  • Zhang B  et al. Mesenchymal stem cells rejuvenate cardiac muscle after ischemic injury. Aging (Albany NY) 11:63-72 (2019). Read more (PubMed: 30613028) »
  • Chen YJ  et al. Inhibition of the potassium channel Kv1.3 reduces infarction and inflammation in ischemic stroke. Ann Clin Transl Neurol 5:147-161 (2018). Read more (PubMed: 29468176) »
See all 17 Publications for this product

Customer reviews and Q&As

Filter by Ratings

1-7 of 7 Abreviews

TUNEL staining on Lewis Lung Carcinoma cells in vitro

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Worked very well on mouse Lewis Lung Carcinoma cells grown in vitro.

Dr. Louise Reynolds

Verified customer

Submitted Apr 05 2019

Excellent on human tumor tissue

Excellent Excellent 5/5 (Ease of Use)
Abreviews
This kit is very good when I used on human tumor sections. Just follow the manual and you will get good results. Also, if you want do double fluorescent staining for other antigen, you can use heat induced antigen retrieve too.

Dr. Hongwei Shao

Verified customer

Submitted Dec 19 2018

Abreviews
It works on quail tissue.
It is a long procedure, it fades relatively fast.

Abcam user community

Verified customer

Submitted Nov 28 2018

Flow Cyt

Average Average 3/5 (Ease of Use)
Abreviews
I tried to used (ab66110) For the Flow Cyt after fixing the cells it dose not give any differences between the positive and negative samples (dose not work)

Abcam user community

Verified customer

Submitted Nov 15 2018

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

Abcam user community

Verified customer

Submitted Sep 08 2017

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.

Abcam user community

Verified customer

Submitted Apr 07 2016

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.

Mr. James Gahan

Verified customer

Submitted Oct 06 2014

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

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