Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Protocol
  • Published:

The JAM Test and its daughter P-JAM: simple tests of DNA fragmentation to measure cell death and stasis

Abstract

Cytotoxic T lymphocytes, and other death-inducing agents, have at least two different ways of killing their targets: drilling holes in the target cell membrane, or triggering the targets to commit suicide. The JAM Test is a method that measures the DNA fragmentation that accompanies cell suicide. We label target cells with radioactive DNA-precursor nucleotides and harvest them onto fiberglass filters, which trap large pieces of DNA but pass smaller fragments of apoptotic cells. As a general measure of apoptosis, the JAM Test described here is faster (can be completed in 4 h [or less if labeling is done the night before]), more quantitative, easier, more sensitive, more flexible and cheaper than most other current assays of apoptosis. The P-JAM, also discussed, additionally allows for assessment of death in cells that don't fragment their DNA, and allows for assays of agents that induce cell stasis rather than death.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Overview of setting up and running the JAM Test and P-JAM.
Figure 2: Layout of 96-well plate.
Figure 3: Comparison of CTL-mediated killing over time.
Figure 4: A 4-h P-JAM detects killing of adherent tumor cells that a 4-h JAM Test does not detect.

Similar content being viewed by others

References

  1. Matzinger, P. The JAM test. A simple assay for DNA fragmentation and cell death. J. Immunol. Methods 145, 185–192 (1991).

    Article  CAS  Google Scholar 

  2. D'Adamio, L. et al. Negative selection of thymocytes. A novel polymerase chain reaction-based molecular analysis detects requirements for macromolecular synthesis. J. Immunol. 149, 3550–3553 (1992).

    CAS  PubMed  Google Scholar 

  3. Böhm, C. et al. A modification of the JAM test is necessary for a correct determination of apoptosis induced by FasL+ adherent tumor cells. J. Immunol. Methods 217, 71–78 (1998).

    Article  Google Scholar 

  4. Ayres, F.M. et al. A comparative study of the JAM test and 51Cr-release assay to assess the cytotoxicity of dendritic cells on hematopoietic tumor cells. Immunol. Invest. 32, 219–227 (2003).

    Article  CAS  Google Scholar 

  5. van Genderen, H. et al. In vitro measurement of cell death with the annexin A5 affinity assay. Nat. Protocols 1 (doi:10.1038/nprot.2006.55) (2006).

  6. Hoves, S. et al. The JAM-assay: optimized conditions to determine death-receptor-mediated apoptosis. Methods 31, 127–134 (2003).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We would like to thank the numerous people who, over the years, asked questions, sent suggestions, and shared their successes and failures with the JAM Test. This research was supported by the Intramural Research Program of the NIAID, NIH.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Polly Matzinger.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Usharauli, D., Perez-Diez, A. & Matzinger, P. The JAM Test and its daughter P-JAM: simple tests of DNA fragmentation to measure cell death and stasis. Nat Protoc 1, 672–682 (2006). https://doi.org/10.1038/nprot.2006.107

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nprot.2006.107

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing