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Related Experiment Videos

Polysome isolation of sepharose column chromatography.

W H Eschenfeldt, R J Patterson

    Preparative Biochemistry
    |January 1, 1975
    PubMed
    Summary

    Researchers purified polysomes from mouse myeloma cells using Sepharose gel filtration. This method effectively removed smaller cellular components and preserved polysome integrity during long-term storage at -85°C.

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    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Cell Biology

    Background:

    • Polysomes are crucial for protein synthesis.
    • Efficient purification of intact polysomes is essential for biochemical studies.
    • Previous methods may not fully remove contaminants or preserve polysome structure.

    Purpose of the Study:

    • To evaluate the efficacy of Sepharose gel filtration for polysome purification.
    • To assess the impact of different Sepharose column sizes (6B, 4B, 2B) on purification.
    • To determine the stability of purified polysomes under prolonged cold storage.

    Main Methods:

    • Purification of polysomes from mouse myeloma MOPC-21 cells.
    • Employing gel filtration chromatography with Sepharose 6B, 4B, and 2B columns.
    • Analysis of purified fractions for polysomal integrity and removal of contaminants.

    Main Results:

    • All three Sepharose columns effectively removed intracellular material smaller than 40 S subunits.
    • Sepharose 4B and 2B columns significantly reduced the presence of subunits and monosomes.
    • Purified polysomes maintained structural integrity after storage at -85°C for at least nine weeks.

    Conclusions:

    • Sepharose gel filtration is an effective method for purifying intact polysomes.
    • Smaller pore-sized Sepharose columns (4B, 2B) offer superior removal of contaminants.
    • Purified polysomes are stable for extended periods when stored at low temperatures, facilitating further research.

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