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Related Experiment Videos

An improved europium release assay for complement-mediated cytolysis.

J Cui1, J C Bystryn

  • 1Department of Dermatology, New York University School of Medicine, NY 10016.

Journal of Immunological Methods
|February 14, 1992
PubMed
Summary

A novel europium release assay offers a sensitive and rapid method for measuring complement-mediated cytolysis. This improved assay detects cell death more effectively than traditional methods, simplifying experimental procedures.

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Area of Science:

  • Immunology
  • Biochemistry
  • Cell Biology

Background:

  • Complement-mediated cytolysis is crucial in immune responses.
  • Existing assays for cytolysis, such as 51Cr release and trypan blue exclusion, have limitations in sensitivity and speed.
  • There is a need for improved, quantitative methods to assess complement-mediated cell death.

Purpose of the Study:

  • To develop and validate an improved assay for measuring complement-mediated cytolysis.
  • To enhance the sensitivity and efficiency of cytolysis detection.
  • To provide a simpler and more quantitative alternative to conventional assays.

Main Methods:

  • Target cells were labeled with europium complexed to diethylenetriaminopentaacetate (Eu-DTPA).
  • Cytolysis was induced by antibody plus complement, leading to the release of the Eu-DTPA complex.

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  • Released europium was quantified using a time-resolved fluorometer after chelation with 2-naphthoyltrifluoroacetone (2-NTA).

    • Main Results:

    • The europium release assay demonstrated high reproducibility.
    • This assay was five times more sensitive than 51Cr release and three times more sensitive than trypan blue exclusion for measuring complement-mediated cytolysis.
    • Europium labeling allows for batch processing and frozen storage of target cells, simplifying the assay workflow.

    Conclusions:

    • The europium release assay is a simple, quantitative, sensitive, and rapid method for measuring complement-mediated cytolysis.
    • This assay offers significant advantages over conventional methods in terms of sensitivity and speed.
    • The assay's robustness and ease of use make it a valuable tool for immunological research.