Proteins from metaphase chromosomes treated with fluorochromes

Area of Science:

• Molecular Biology
• Cell Biology
• Biochemistry

Background:

• Isolation and purification procedures can alter the biochemical composition of organelles.
• Fluorochromes are commonly used in flow cytometry but their effect on chromosome protein composition is not well understood.

Purpose of the Study:

• To investigate the impact of fluorochromes and staining conditions on the protein profiles of isolated metaphase chromosomes.
• To determine if biochemical analysis of flow cytometry-purified chromosomes is feasible.

Main Methods:

• Metaphase chromosomes were exposed to common fluorochromes and staining conditions used in flow cytometry.
• Protein profiles were analyzed using polyacrylamide gel electrophoresis (PAGE).
• Electropherograms were compared to control samples.

Main Results:

• PAGE analysis showed that fluorochrome exposure did not alter the protein profiles of metaphase chromosomes.
• Electropherograms of treated and control chromosomes were indistinguishable.

Conclusions:

• Fluorochromes and associated staining conditions do not significantly affect the protein composition of isolated chromosomes.
• Biochemical analysis of individual chromosomes sorted by flow cytometry is now a validated approach.