For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Customized Products & Partnerships
    Customized Products & Partnerships

    Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs.

    Customized products

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

  1. Link

    tunel-assay-kit-fitc-ab66108.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Kits/ Lysates/ Other Kits Apoptosis Kits DNA Fragmentation TUNEL Assay Kits
Share by email

TUNEL Assay Kit - FITC (ab66108)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (5)References (32)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Shipping info

Promotion Information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

TUNEL assay staining
  • Functional assays: In situ Direct DNA Fragmentation (TUNEL) Assay Kit (ab66108)
  • Functional assays: In situ Direct DNA Fragmentation (TUNEL) Assay Kit (ab66108)
  • Functional assays: In situ Direct DNA Fragmentation (TUNEL) Assay Kit (ab66108)

Key features and details

  • Assay type: Direct
  • Detection method: Fluorescent
  • Platform: Flow cytometer, Fluorescence microscope
  • Assay time: 2 hr 30 min
  • Sample type: Adherent cells, Suspension cells

You may also be interested in

ELISA
Product image
BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
Assay
Product image
Annexin V-FITC Apoptosis Staining / Detection Kit (ab14085)
Assay
Product image
Caspase-3, Caspase-8 and Caspase-9 Multiplex Activity Assay Kit (Fluorometric) (ab219915)

View more associated products

Overview

  • Product name

    TUNEL Assay Kit - FITC
    See all DNA fragmentation kits
  • Detection method

    Fluorescent
  • Sample type

    Adherent cells, Suspension cells
  • Assay type

    Direct
  • Assay time

    2h 30m
  • Product overview

    TUNEL Assay Kit - FITC ab66108 provides complete components, including positive and negative control cells, for detecting DNA fragmentation by fluorescence microscopy or flow cytometry.


    The TUNEL assay is used to detect DNA fragmentation, such as in apoptosis. It uses terminal deoxynucleotidyl transferase (TdT) to catalyze the incorporation of deoxynucleotides at the free 3'-hydroxyl ends of fragmented DNA. The deoxynucleotides are then labeled in a variety of ways for detection of the degree of DNA fragmentation.


    The TUNEL assay protocol in this kit uses the deoxynucleotide fluorescein-12-dUTP. DNA which has been labeled with fluorescein can then be analyzed by flow cytometry or fluoresence microscopy with Ex/Em 488/520 nm. Propidium iodide is also included in this kit as a counterstain with Ex/Em 488/623 nm.


    TUNEL assay protocol summary:
    - fix cells with formaldehyde for 15 min on ice
    - wash with PBS
    - add ice-cold 70% ethanol and incubate for 30 min
    - pellet cells and resuspend in wash buffer and wash again
    - pellet cells and resuspend in staining solution and incubate for 60 min at 37ºC
    - add rinse buffer, pellet cells and discard supernatant, and rinse again
    - resuspend cells in propidium iodide/RNAse A solution and incubate for 30 min at room temp
    - analyze with flow cytometer or fluoresence microscope

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called K402 ApoDIRECT DNA Fragmentation Assay Kit. K402-50 is the same size as the 50 test size of ab66108.

    This kit is FITC-labeled (Ex/Em = 495/519nm). It was previously called TUNEL Assay Kit - In situ Direct DNA Fragmentation.

    To use BrdU-Red (Ex/Em = 488/576nm) as a label, we recommend TUNEL Assay Kit - BrdU-Red (ab66110).

    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.


    How other researchers have used FITC TUNEL Assay Kit ab66108

    This TUNEL assay kit has been used in publications in a variety of sample types, including:
    - Human: HUVEC cell cultures1, AGS gastric carcinoma cells by imaging2, gastric tumor cell xenograft cells by flow cytometry3, MDA-MB-231 breast cancer xenograft tissue sections by imaging4, neural blastoma cell cultures by flow cytometry5, SH-SY5Y cells by flow cytometry6, A549 cells by flow cytometry7, Huh7 and HepG2 cell cultures by imaging8
    - Mouse: liver tissue by imaging9, cultured neural stem cells by imaging10
    - Rat: kidney tissue sections by imaging11

    References: 1 - De Felice F et al 2019, 2 - Li C et al 2019, 3 - Lau WM et al 2018, 4 - Chung SJ et al 2017, 5 - Sobham PK et al 2017, 6 - Albarran L et al 2016, 7 - Lamb SA et al 2014, 8 - Fu B et al 2016, 9 - Azam F et al 2018, 10 - Voloboueva LA et al 2017, 10 - Sun X et al 2015, 11 - Chen J et al 2015

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Flow cytometer, Fluorescence microscope

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 50 tests
    FITC-dUTP Orange 1 x 0.4ml
    Negative Control Cells Neutral 1 x 5ml
    PI/RNase Staining Buffer Amber bottle 1 x 25ml
    Positive Control Cells Brown 1 x 5ml
    Reaction Buffer Green 1 x 500µl
    Rinse Buffer Red 1 x 100ml
    TdT Enzymes 1 x 38µl
    Wash Buffer Blue 1 x 100ml
  • Research areas

    • Kits/ Lysates/ Other
    • Kits
    • Apoptosis Kits
    • DNA Fragmentation
    • TUNEL Assay Kits
    • Kits/ Lysates/ Other
    • Kits
    • Molecular Biology Kits
    • DNA Isolation Kits
    • Kits/ Lysates/ Other
    • Kits
    • Apoptosis Kits
    • DNA Fragmentation
    • DNA ladder
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Cell Damage
  • Relevance

    Internucleosomal DNA fragmentation is a hallmark of apoptosis in mammalian cells.

Associated products

  • Assay kits

    • TUNEL Assay Kit - HRP-DAB (ab206386)
    • TUNEL Assay Kit - BrdU-Red (ab66110)
  • Related Buffer

    • Z-VAD(OH)-FMK, Irreversible general caspase inhibitor (ab120382)
  • Related Products

    • FCCP, mitochondrial oxidative phosphorylation uncoupler (ab120081)
    • Cycloheximide, Protein synthesis inhibitor (ab120093)
    • SB431542, ALK inhibitor (ab120163)
    • Z-VAD(OMe)-FMK, Cell permeable, irreversible pan-caspase inhibitor (ab120487)
    • Doxorubicin hydrochloride, Topoisomerase II inhibitor (ab120629)
    • Dorsomorphin (Compound C), AMP-kinase inhibitor (ab120843)
    • Calcein AM, fluorescent dye for cell viability (ab141420)

Images

  • TUNEL assay staining
    TUNEL assay stainingVoloboueva LA et al. J Neurosci 37:3072-3084 (2017) Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Voloboueva et al used TUNEL assay ab66108 to examine the effect of miR-210 inhibition on mitochondrial function and protection against apoptosis.

    The reduced number of TUNEL+ve green cells in panel F indicates that the inhibition of miR-210 reduces the degree of apoptosis  in cells treated with media preconditioned (CM) by proinflammatory activated microglia.

    Green is TUNEL staining. Red is immunostaining of DCX protein. Representative images are shown.

     

     

  • Functional assays: In situ Direct DNA Fragmentation (TUNEL) Assay Kit (ab66108)
    Functional assays: In situ Direct DNA Fragmentation (TUNEL) Assay Kit (ab66108)

    Control RAW 264.7 cells.

  • Functional assays: In situ Direct DNA Fragmentation (TUNEL) Assay Kit (ab66108)
    Functional assays: In situ Direct DNA Fragmentation (TUNEL) Assay Kit (ab66108)

    RAW 264.7 cells treated with 2 μM camptothecin (ab120115) for 24 hours prior to staining.

  • Functional assays: In situ Direct DNA Fragmentation (TUNEL) Assay Kit (ab66108)
    Functional assays: In situ Direct DNA Fragmentation (TUNEL) Assay Kit (ab66108)

    RAW 264.7 cells treated with 10 μM camptothecin (ab120115) for 24 hours prior to staining.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (32)

Publishing research using ab66108? Please let us know so that we can cite the reference in this datasheet.

ab66108 has been referenced in 32 publications.

  • Dolicka D  et al. Tristetraprolin Promotes Hepatic Inflammation and Tumor Initiation but Restrains Cancer Progression to Malignancy. Cell Mol Gastroenterol Hepatol 11:597-621 (2021). PubMed: 32987153
  • D'Souza AM  et al. Small Molecule Cjoc42 Improves Chemo-Sensitivity and Increases Levels of Tumor Suppressor Proteins in Hepatoblastoma Cells and in Mice by Inhibiting Oncogene Gankyrin. Front Pharmacol 12:580722 (2021). PubMed: 33746747
  • Yang L  et al. Azeliragon ameliorates Alzheimer's disease via the Janus tyrosine kinase and signal transducer and activator of transcription signaling pathway. Clinics (Sao Paulo) 76:e2348 (2021). PubMed: 33681944
  • Gu R & Zhang Q Effects of low-dose bufalin combined with hydroxycamptothecin on human castration-resistant prostate cancer xenografts in nude mice. Exp Ther Med 22:1015 (2021). PubMed: 34373701
  • Liaudanskaya V  et al. Modeling Controlled Cortical Impact Injury in 3D Brain-Like Tissue Cultures. Adv Healthc Mater 9:e2000122 (2020). PubMed: 32406202
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-5 of 5 Abreviews or Q&A

Question

Customer kindly contacted us regarding ab66108. After storing samples at -20C in ethanol, he is not seeing a pellet after resuspension in step 2 b). He has about 3 million cells harvested initially. Should he be able to see a pellet at this point?

Read More

Abcam community

Verified customer

Asked on Jan 25 2013

Answer


The laboratory responded to your inquiry regarding visibility of a pellet after adding the washing buffer. You should be able to see at least a mini-speck of the pellet even after the wash step. One easy thing to do would be to take an aliquot of the cells in the second step of washing and look at it under the microscope. If you are able to see the cells, then they are there.

Read More

Abcam Scientific Support

Answered on Jan 25 2013

Question

in situ direct DNA fragmantation KIT protocol mentioned cells not tissue, my sample are slides of paraffin embedded tissue can this kit modified to work on tissue slides and examined by fluorescent microscope.

Read More

Abcam community

Verified customer

Asked on Jan 03 2013

Answer

We have not tried this protocol on tissue sections, but here is a protocol that the lab has found from another which can be modified to fit into this procedure. A. Tissue Section Preparations: The protocol describes the preparation of formalin-fixed, paraffin-embedded tissue section mounted on glass slides. For information on fixing and embedding techniques, see Ben-Sasson et al., (Methods Cell. Biol. 46:29-39, 1995). Most steps are performed in Coplin Jars. Note: If using fresh-frozen tissue sections, proceed directly to step 7. 1. Remove paraffin by immersing slides in a Coplin jar containing fresh xylene. Incubate at room temperature for 5 minutes. 2. Repeat in a second Coplin jar containing fresh xylene. 3. Immerse the slides in a Coplin Jar containing 100% ethanol and incubate at room temperature for 5 min. 4. Rehydrate the slides by sequential 3-min, room temperature incubations in Coplin jars containing: · 100% ethanol · 95% ethanol · 85% ethanol · 70% ethanol · 50% ethanol 5. Immerse the slides in a Coplin jar containing 0.85% NaCl and incubate at room temperature for 5 min. 6. Immerse the slides in a Coplin jar containing PBS and incubate at room temperature for 5 minutes. 7. Fix the slides by immersing them in a Coplin jar containing fresh 4% formaldehyde/PBS, and incubate at room temperature for 15 min. 8. Wash the slides by immersing them in a Coplin jar containing PBS, and incubate at room temp. for 5 min. 9. Transfer to another Coplin jar containing PBS, and incubate at room temperature for 5 min. 10. Allow the liquid to drain thoroughly and place slides on a flat surface. 11. Prepare 20 µg/ml of Proteinase K Solution (combine 2 µl of 10 mg/ml Protease K and 998 ml of 100 mM Tris-HCl, pH 8.0, 50 mM EDTA) and cover each section with 100 µl of it. Incubate at room temperature for 5 min. 12. Immerse the slides in Coplin jar containing PBS, and incubate at room temperature for 5 min. 13. Transfer the slides to a Coplin jar containing 4% formaldehyde/PBS and incubate at room temperature for 5 minutes. 14. Wash the slides by immersion in Coplin jar containing PBS, and incubate at room temperature for 5 min. B. Detection by Fluorescence Microscopy: 1. Remove slides from PBS and tap gently to remove excess liquid. Cover the cells in 100 µl of Wash buffer (blue cap). 2. Use forceps, gently place a piece of plastic coverslip on top of the cells to evenly spread the liquid, incubate for 5 min. Remove plastic coverslip and gently tap the slides to remove excess liquid. 3. Repeat step 2. Carefully blot dry around the edges with tissue paper. 4. Gently place 50 µl of the DNA Labeling Solution (prepared as in Section IIIB, Step 4) on the cells. 5. Use forceps, gently place a piece of plastic coverslip on top of the cells to evenly spread the liquid. 6. Place the slides in a dark, humidified 37°C incubator for 60 minutes. Note: Ensure high humidity by placing wet paper towels in the bottom of the dry incubator. Antibody Solution 1 assay 10 assays Anti-BrdU-FITC Antibody (orange cap) 5 µl 50 µl Rinse Buffer (red cap) 95 µl 950 µl 7. Using forceps, remove the plastic coverslips. Rinse the slides to a fresh Coplin jar filled with PBS for 5 min. 8. Repeat step 7. Carefully blot dry around the edges with tissue paper. 9. Place 100 µl of the Antibody Solution (Prepared as in Section IIIB, step 8). 10. Use forceps, gently place a piece of plastic coverslip on top of the cells to evenly spread the liquid. 11. Incubate the cells with the antibody solution in a humidified incubator for 30 min at room temperature. 12. Carefully remove the solution from slides. Add 100 µl of Propidium Iodide/Rnase A solution (amber bottle). 13. Use forceps; gently place a piece of plastic coverslip on top of the cells to evenly spread the liquid. 14. Incubate the slides in the dark in a humidified incubator for 30 min at room temperature. 15. Wash the cells by transferring the slides to a fresh Coplin jar filled with ddH2O and incubate at room temperature for 5 min. 16. Repeat Step 15. 17. [Optional] Add a drop of anti-Fade solution and cover the treated portion of the slide with a glass coverslip. 18. [Optional] Seal the edges of the coverslip with rubber cement or clear nail polish. 19. View slides as soon as possible using FITC and rhodamine filters. Apoptotic cells will exhibit strong nuclear green fluorescence. All cells should be stained with PI and exhibit strong red counter staining.

Read More

Abcam Scientific Support

Answered on Jan 03 2013

Question

dear technical team i had a previous inquiry with number:CCE 4458713 about the kit of direct DNA fragmantation assay kit, i need you to provide me with the detailed protocol (technique) of this kit application on paraffin sections on postive charged slides. with best regards.

Read More

Abcam community

Verified customer

Asked on Dec 21 2012

Answer

Thank you for your inquiry and your patience while we looked into this further.

For ab66108, the kit components are optimized for usage on cells. If using tissue sections, you will not be able to get a full 50 assays from the kit because you will run short of reaction buffer.
Here’s the protocol for using ab66108 for paraffin embedded tissue sections:
DEPARAFFINIZATION AND REHYDRATION
1. Immerse slides in xylene (or xylene substitute) for 5 minutes at room temperature. Repeat using fresh xylene for second 5 minute incubation.
2. Immerse slides in 100% ethanol for 5 minutes at room temperature. Repeat using fresh 100% ethanol for second 5 minute incubation.
3. Immerse slides in 90% ethanol for 3 minutes at room temperature.
4. Immerse slides in 80% ethanol for 3 minutes at room temperature.
5. Immerse slides in 70% ethanol for 3 minutes at room temperature.
6. Immerse slides briefly into 1X PBS and carefully dry the glass slide around the specimen.
Do not let the tissue specimen dry out during or between any step!
(If necessary, cover or immerse the specimen in 1X PBS to keep hydrated)
** At this point it may be helpful to encircle the specimen using a waxed pen or a hydrophobic marker.
PERMEABILIZATION OF SPECIMEN
1. Dilute enough Proteinase K (STOCK: 2 mg/ml in 10 mM Tris pH 8) needed 1:100 in 10 mM Tris pH 8.
2. Cover the entire specimen with 100 μl diluted proteinase K. Incubate at room temperature for 20 minutes. DO NOT OVER INCUBATE.
3. Rinse slide with 1X PBS.
4. Gently tap off excess liquid and carefully dry the glass slide around the specimen.
EQUILIBRATION AND Staining REACTION
1. Dilute Reaction Buffer (green cap) as needed 1:5 with dH2O.
(NOTE: This buffer does NOT contain nucleotides or enzyme).
2. Cover the entire specimen with 100 μl of the diluted Reaction Buffer. Incubate at room temperature for 10 to 30 minutes while preparing the staining reaction mixture.
3. For making staining solution, follow K402-100 kit protocol.
4. Carefully blot the Reaction Buffer from the specimen, taking care not to
touch the specimen.
5. Immediately apply 50 μl of staining solution onto each specimen.
** The use of a cover slip at this point assures even distribution of the reaction mixture and prevents loss due to evaporation during incubation. **
6. Cover the specimen with a piece of Parafilm cut slightly larger than the specimen. HINT: Folding up one corner of the Parafilm cover slip will aid in its application and removal.
7. Place slides in a humid chamber and incubate at 37C for 1 to 1.5 hours Note: incubation times at 37C may need to be adjusted to longer or shorter periods depending on the characteristics of the tissue supplied by the researcher..
8. Remove Parafilm cover slip and rinse slide with 1X PBS.
9. Repeat the rinsing step.
10. Gently tap off excess liquid and carefully dry the glass around the specimen.
11. Dilute PI/RNase staining buffer 1:5 in PBS & apply 50 ul of diluted Propidium Iodide/RNase A Solution (amber bottle) onto the specimen.
12. Incubate in the dark for 30 min at room temperature.
13. Analyze by fluorescence microscopy (apoptotic cells show green staining over an orange-red PI counter-staining)




I hope this information helps. Please contact us with any other questions.

Read More

Abcam Scientific Support

Answered on Dec 21 2012

Question


Inquiry: dear technical team greatly appreciating your help, your kit in the protocol deal with sample as collection of cells . in my work i would like to preserve the tissue in its architecture, fixed in neutral buffered formaldehyde, prepared in paraffin sections,on positive silinized slides. can your kit work on such sample with modification as addition of protinase K. please inform me as soon as possible

Read More

Abcam community

Verified customer

Asked on Dec 13 2012

Answer

Thank you for contacting us. This kit will work fine with the samples you describe.

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

Read More

Abcam Scientific Support

Answered on Dec 13 2012

Question

Dear technical team,   Our customer would like to purchase TUNEL assay product so, I produced ab66108 and ab66110.   It described that sample types are cells and tissue on the page 6 of Assay Kits brochure but,   I cann't find the protocol for tissue on the website.   1. Are those available for tissue samples?   2. If yes, send me the specific protocol please.     Thank you.   Best regards,

Read More

Abcam community

Verified customer

Asked on Sep 28 2011

Answer

Thank you for your enquiry and your interest in our products. The two kits (ab66108 and ab66110) are mainly used for cells analyzed by flow cytometry. Currently, we do not have specific kits for tissue samples (for DNA Fragmentation) but I would advise you and your customers to check our catalogue regularly since we add and publish new products every day. Apologies! If you need any further assistance in the future, please do not hesitate to contact me.

Read More

Abcam Scientific Support

Answered on Sep 28 2011

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Get resources and offers direct to your inbox Sign up
A-Z by research area
  • Cancer
  • Cardiovascular
  • Cell biology
  • Developmental biology
  • Epigenetics & Nuclear signaling
  • Immunology
  • Metabolism
  • Microbiology
  • Neuroscience
  • Signal transduction
  • Stem cells
A-Z by product type
  • Primary antibodies
  • Secondary antibodies
  • Biochemicals
  • Isotype controls
  • Flow cytometry multi-color selector
  • Kits
  • Loading controls
  • Lysates
  • Peptides
  • Proteins
  • Slides
  • Tags and cell markers
  • Tools & Reagents
Help & support
  • Support
  • Make an Inquiry
  • Protocols & troubleshooting
  • Placing an order
  • RabMAb products
  • Biochemical product FAQs
  • Training
  • Browse by Target
Company
  • Corporate site
  • Investor relations
  • Company news
  • Careers
  • About us
  • Blog
Events
  • Tradeshows
  • Conferences
International websites
  • abcam.cn
  • abcam.co.jp

Join with us

  • LinkedIn
  • facebook
  • Twitter
  • YouTube
  • Terms of sale
  • Website terms of use
  • Cookie policy
  • Privacy policy
  • Legal
  • Modern slavery statement
© 1998-2022 Abcam plc. All rights reserved.