Mammalian Cell Lysis Protocols

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Mammalian Cell Lysis Protocols
Cell lysis is needed for a number of laboratory techniques. (Jupiterimages/Comstock/Getty Images)

Cell lysis is the breaking open of cells, while causing as little damage to the proteins, organelles, DNA and other macromolecules as possible. Cell lysis is used for a number of cell and molecular biology techniques, including Western blotting, analysis of proteins, Southern blotting and study of DNA sequence. The molecule or organelle under study will determine the method used to lyse the cell.

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Mechanical

There are a number of mechanical methods to lyse cells including blenders, ultrasound and homogenizers. This method relies on blades to grind and displace the tissues and cells. It often is used for cells that are difficult to lyse, such as microorganisms. However, complex tissues, such as liver or kidney, may require mechanical lysis. Mechanical lysis often results in a viscous solution that is difficult to clarify. The heat generated may denature proteins of interest.

Buffers

Detergents often are used to lyse mammalian cells. This method is milder by disrupting the cells' membranes. The detergent selected will depend on what techniques the cell lysate is to be used for and the cells that need to be lysed. The cells and buffers often are kept cold if you are studying proteins to prevent their denaturation. Other considerations are the pH and salt concentrations and if inhibitors are needed to prevent denaturation of proteins.

Freeze Thaw

Cells can be frozen and thawed and, over a number of cycles, the cell will lyse. This method causes the cell to swell and ice crystals form, which causes the cell membrane to break and release the cell's contents. A study by Haifeng Yu and colleagues for Bio-Rad, in 2006, showed that the addition of a freeze-thaw cycle with sonication, in mammalian cell types, improved the sensitivity for some proteins compared to just sonication.

Liquid Homogenization

Cells are forced through a narrow space, which breaks down the cell membrane. A number of different types of homogenizers are available, though the equipment can be expensive. The number of times that the cells have to pass through depends on the cells and equipment used. Yield, however, can be variable.

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