In HBV research, HBV DNA Southern blot preparation is done by lysis of cells and viral particles. In that case, it used cold lysis buffer contained 10mM HEPES and 0.01X NP-40. What are the functions of those two compounds in lysis buffer?
Hello Kamindu Gayashan
HEPES is a zwitterionic buffering reagent which functions over a pH range of 6.5 to 8.5. It maintains the ion concentration of a solution at a level that's ideal for a certain purpose. The pH level of the solution is maintained in this manner. Its presence in the lysis buffer will have a significant impact on protein stability as well as on biological performance.
NP-40 is a milder, nonionic detergent that is frequently used as a component of cell lysis buffers intended to extract and solubilize proteins. It is good at solubilizing membrane proteins and for isolating cytoplasmic proteins. Proteins retain their native state in the presence of this detergent and protein-protein interactions can be preserved. NP-40 will not lyse nuclear membranes.
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