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Related Experiment Videos

A rapid procedure for isolating hemopoietic cell nuclei.

C M Bunce1, J A Thick, J M Lord

  • 1Department of Immunology, University of Birmingham, United Kingdom.

Analytical Biochemistry
|November 15, 1988
PubMed
Summary

A rapid new method efficiently isolates pure cell nuclei using freezing, thawing, and sucrose centrifugation. This technique preserves nuclear integrity and minimizes contamination for accurate biochemical analysis.

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Molecular Biology

Background:

  • Isolating pure cell nuclei is crucial for studying nuclear function and composition.
  • Traditional methods can be time-consuming and may lead to loss or modification of nuclear components.

Purpose of the Study:

  • To develop and validate a rapid, efficient method for isolating intact cell nuclei.
  • To minimize loss of nuclear proteins and metabolites during isolation.

Main Methods:

  • Cells were frozen and thawed in 2% Tween 40.
  • Gentle homogenization released nuclei.
  • Immediate microcentrifugation through 50% sucrose purified nuclei.

Main Results:

  • Purified nuclei were obtained within 3 minutes.

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  • Yields ranged from 78-95% from human hemopoietic cells.
  • Electron microscopy confirmed 89% intact nuclei with appropriate morphology and low organelle contamination (0.5-5.5%).

    • Conclusions:

    • The described method provides rapid and high-yield isolation of pure, intact cell nuclei.
    • This technique minimizes nuclear component loss and biochemical modification, making it valuable for nuclear protein and metabolite studies.