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Microsome Preparation Protocol

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    Methods

    • Scientists use a variety of methods to break open cells: sonicators, freeze-thaw cycles, homogenizers, detergents and for the larger samples, blenders. They centrifuge this slurry at slow speed to pellet the bigger pieces -- nuclei and unbroken cells -- and this pellet is discarded. The remaining suspension is centrifuged at high speed to pellet the microsomes -- these are resuspended in a buffer for use.

    Considerations

    • The scientist should consider a number of points before starting. First would be the cell type -- for instance liver should be perfused to remove contaminating blood. Second, molecules to be studied may require a particular temperature for stability. Third, the microsomes can be separated into different sizes, therefore the centrifuge speed can be critical for obtaining the desired fraction.

    Uses

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