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Monitoring early cellular responses in apoptosis is aided by the mitochondrial membrane protein-specific monoclonal

S K Koester1, P Roth, W R Mikulka

  • 1Department of Scientific Research, Coulter Technology Center, Miami, Florida 33196, USA.

Cytometry
|January 24, 1998
PubMed

Insights

The APO2.7 antibody effectively monitors early apoptosis by detecting mitochondrial changes in cells. This antibody provides a reliable method for identifying apoptotic cells, even before significant DNA fragmentation occurs.

Area of Science:

  • Cell Biology
  • Immunology
  • Biochemistry

Background:

  • Apoptosis, or programmed cell death, is a crucial biological process.
  • Monitoring early apoptotic events is vital for understanding cellular responses to stimuli.
  • Existing methods for detecting apoptosis have limitations in sensitivity and timing.

Purpose of the Study:

  • To evaluate the efficacy of the APO2.7 monoclonal antibody in detecting early apoptotic responses.
  • To compare APO2.7 antibody performance with other apoptosis markers in anti-CD95-induced Jurkat cells.
  • To determine the optimal conditions for using APO2.7 for apoptosis monitoring.

Main Methods:

  • Jurkat cells were induced to undergo apoptosis using anti-CD95 (7C11).
  • Cells were analyzed at various time points (1.5-18 h) using flow cytometry.
  • APO2.7 antibody staining was performed on both unprocessed and processed (permeabilized) cells.
  • Apoptosis markers including light-scatter changes, cell permeability (Trypan blue), tubulin, DNA fragmentation, and phosphatidylserine exposure (annexin V) were assessed.

Main Results:

  • Light-scatter changes indicative of apoptosis were observed after 3 hours.
  • Cell permeability and tubulin changes were detected after 6 hours in unprocessed cells.
  • APO2.7 staining showed a significant increase in processed cells by 1.5 hours, and in unprocessed cells by 6 hours.
  • APO2.7 staining in processed cells correlated well with annexin V detection from 3-18 hours.

Conclusions:

  • The APO2.7 antibody is a valuable tool for detecting early apoptotic events.
  • APO2.7 can identify apoptotic cells by targeting mitochondrial membrane proteins.
  • The antibody's utility is enhanced in processed cells, allowing for earlier detection of apoptosis.

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