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Related Experiment Videos

Different intermediate-sized filaments distinguished by immunofluorescence microscopy.

W W Franke, E Schmid, M Osborn

    Proceedings of the National Academy of Sciences of the United States of America
    |October 1, 1978
    PubMed
    Summary
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    Researchers identified vimentin, a 57,000 MW protein in mouse cells, distinguishing intermediate filaments. Immunofluorescence microscopy revealed vimentin in mesenchymal cells, while prekeratin marked epithelial cells, differentiating filament types.

    Area of Science:

    • Cell Biology
    • Biochemistry
    • Immunology

    Background:

    • Intermediate-sized filaments are crucial cytoskeletal components.
    • Distinguishing different intermediate filament types is essential for understanding cellular structure and function.
    • Previous characterization of these filaments lacked specific immunological markers.

    Purpose of the Study:

    • To identify and characterize the major protein of intermediate-sized filaments in mouse 3T3 cells.
    • To develop and utilize antibodies for distinguishing different intermediate filament populations.
    • To investigate the distribution of vimentin and prekeratin in various cell types and tissues.

    Main Methods:

    • Protein purification and molecular weight determination.
    • Production of antibodies against purified vimentin and prekeratin.

    Related Experiment Videos

  • Immunofluorescence microscopy on cultured cells and frozen tissue sections.

    • Main Results:

    • A 57,000 MW protein, named vimentin, was identified as a major component of intermediate filaments.
    • Antibodies against vimentin decorated filaments in non-muscle mesenchymal cells, including transformed cells.
    • Antibodies against prekeratin decorated filaments predominantly in epithelial cells, including tumor-derived cells.
    • Colcemid treatment induced perinuclear aggregates of vimentin filaments.
    • No significant reaction was observed in muscle or brain tissues.

    Conclusions:

    • Intermediate-sized filaments with similar morphology can be immunologically distinguished.
    • Vimentin is a marker for intermediate filaments in non-muscle mesenchymal cells.
    • Prekeratin serves as a marker for intermediate filaments in epithelial cells.
    • Immunological methods provide a powerful tool for classifying intermediate filament systems.