Lysis buffer
A lysis buffer is a buffer solution used for the purpose of breaking open cells for use in molecular biology experiments that analyze the labile macromolecules of the cells (e.g. western blot for protein, or for DNA extraction).
For lysis buffers targeted at protein extraction, protease inhibitors are often included, and in difficult cases may be almost required.
[1] The primary purpose of lysis buffer is isolating the molecules of interest and keeping them in a stable environment.
The important factors to be considered are: pH, ionic strength, usage of detergent, protease inhibitors to prevent proteolytic processes.
Detergents are often categorized as nonionic, anionic, cationic, or zwitterionic, based on their hydrophilic head group feature.
[7] For the longest time, after a detergent-based cell lysis, a buffer exchange and/or dialysis had to be performed to remove the detergent among other hindering compounds to restore native conditions.
The GentleLys buffer employs copolymers instead of detergents, ensuring efficient cell lysis while maintaining the native environment crucial for the correct folding of cellular components, such as proteins.
This gentle approach eliminates the need for harsh chemicals, creating an environment that preserves the native state of cellular proteins.
Consequently, the proteins maintain their structural integrity and functionality, a marked departure from the denaturing effects of detergent-based buffers.
These components include detergents, salts, and enzymes, each playing a specific role in the lysis process.
In the context of cell lysis, detergents act by disrupting the lipid bilayer of the bacterial cell membrane, leading to membrane permeabilization and release of intracellular components, including the target enzyme.
Commonly used detergents in lysing buffers include: a. Triton X-100: a nonionic detergent frequently employed due to its mild and effective membrane-disrupting properties, it solubilizes lipids and membrane proteins, allowing the release of intracellular contents.
Dish soap can be used in a pinch to break down the cell and nuclear membranes, allowing the DNA to be released.
