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Comet Assay Kit (3-well slides) (ab238544)

Reviews (2) Submit a question References (29)
Comet Assay Principle
  • Epifluorescence Microscopy Visualization of DNA Damage.
  • Etoposide Treatment of Jurkat Cells.

Key features and details

  • Detection method: Fluorescent

Overview

  • Product name

    Comet Assay Kit (3-well slides)
    See all DNA damage kits

  • Detection method

    Fluorescent

  • Product overview

    Comet Assay Kit (3-well slides) (ab238544) is a fast and sensitive kit for the measurement of cellular DNA damage.


    The Comet Assay is a single cell gel electrophoresis assay (SCGE) for simple evaluation of cellular DNA damage. It is a convenient way to screen for general DNA damage, regardless of the source or nature of the damage. Kits include Comet Slides, reagents, and a fluorescent dye to visualize cells under an epifluorescence microscope.

  • Notes

    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.

Properties

Associated products

Images

  • Comet Assay Principle
    Comet Assay Principle
    Comet Assay Principle

     

  • Epifluorescence Microscopy Visualization of DNA Damage.
    Epifluorescence Microscopy Visualization of DNA Damage.

    For Data Analysis please see Section 9 in the protocol booklet. 

  • Etoposide Treatment of Jurkat Cells.
    Etoposide Treatment of Jurkat Cells.

    Jurkat cells were untreated (left) or treated (right) with 20 µM Etoposide for 4 hrs before performing Comet Assay (alkaline electrophoresis conditions, 33 V/300 mA for 15 mins).

References (29)

Publishing research using ab238544? Please let us know so that we can cite the reference in this datasheet.

ab238544 has been referenced in 29 publications.

  • von Bülow V  et al. Metabolic reprogramming of hepatocytes by Schistosoma mansoni eggs. JHEP Rep 5:100625 (2023). PubMed: 36590323
  • Zhi W  et al. Short-term starvation synergistically enhances cytotoxicity of Niraparib via Akt/mTOR signaling pathway in ovarian cancer therapy. Cancer Cell Int 22:18 (2022). PubMed: 35016681
  • Keane S  et al. DLG2 impairs dsDNA break repair and maintains genome integrity in neuroblastoma. DNA Repair (Amst) 112:103302 (2022). PubMed: 35217496
  • Kang DY  et al. Antitumor Effects of Natural Bioactive Ursolic Acid in Embryonic Cancer Stem Cells. J Oncol 2022:6737248 (2022). PubMed: 35222644
  • Lorenzoni M  et al. ETS-related gene (ERG) undermines genome stability in mouse prostate progenitors via Gsk3β dependent Nkx3.1 degradation. Cancer Lett 534:215612 (2022). PubMed: 35259458
View all Publications for this product

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1-2 of 2 Abreviews

Ab238544 review comet assay

 Good Average 3/5 (Ease of Use)
Abreviews
Ab238544
After a few preliminary experiments and adaptations, the kit worked well, as we received a typical comet assay image. Additionally, in our case, we quantified those comets and received the expected results in our control samples.
However, we did invest significant time in calibrating and improving the protocol.
7. Sample and Slide Preparation
7.2 We found that upon the first usage, the comet Agarose bottle needed to be heated at 90˚C in a water bath for 20 mins and then aliquoted into 1ml. The advantage of aliquoting is that before any future experiments, all that must be done is to take the desired number of tubes and remelt the Agarose at 90˚C, followed by incubation at 37˚C in a standard heat block.
7.3 base layer preparation. We found that it was much better to add 35-40 µL of Comet Agarose per well and to cover each well with a 16 mm coverslip so that it flattened the first agarose layer. After transferring the slide for 15 mins to 4˚C, and before adding the second layer (cells-agarose layer), the coverslip should be carefully removed.
7.4 cell sample preparation
7.4.2 We used both MDA-MB-231 and A549 adherent cell lines and removed them by trypsinization rather than by using a rubber policeman.
We tried 2 cell concentrations in our experiments; at 1 x 105 cells/mL (as mentioned in the protocol) as well as at 1 x 106 cells/ml, which worked better for our experiments.
7.5 We combined cell samples with Comet Agarose (step 7.2) at a 1/5 ratio (v/v):
A mixture of 7 µl cells with 35µl melted Agarose was immediately transferred onto the top of the Agarose Base Layer (step 7.3) and again was covered by a 16 mm coverslip, ensuring minimum cell layers, which made comet microscopic observation easier.
7.8 Any time the protocol indicated to aspirate the buffer/solution, we took out the liquid by a pipette so that the cells would not be lost.
8. Assay Procedure
In our experiment, we received better results by using TBE electrophoresis rather than Alkaline electrophoresis.
8.1.3 As a previous reviewer reported, we ran the slides at 3v/cm. We found that the running time is cell line dependent and needs to be calibrated. Based on the other reviewer’s recommendation, we ran both cell line slides for a short time (10min), observed the cells under the microscope, and ran only the cell line that required additional time to obtain optimal results.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Sep 16 2022

Electrophoresis conditions need to be optimized, but otherwise working well

 Good Below average 2/5 (Ease of Use)
Abreviews
The method and reagents work well, but the protocol could use some optimization.

I tried several double-strand break generating methods (irradiation, drugs), but I was never able to detect DNA tails after 15 min of electrophoresis in TBE at 1 V/ cm, as described in the protocol.
To see tails, I had to run it at 3 V/ cm for 20 min. Also, it is possible to return the slides to the electrophoresis even after the staining is completed, and run it for longer if needed. This should be mentioned.

The provided agar does not have to be heated to 95C as stated in the protocol as it liquifies at much lower temperatures (I used 70C). This should be mentioned since many waterbaths cannot go to 95C.

These drawbacks aside, I can recommend the product.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 24 2020

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