Dounce homogenization of HeLa suspension cultre - (Jan/27/2010 )
I wanna obtain translationally active lysate from 2 or more litres of HeLa cells in suspension culture, but I don't know what size (2ml or more) of Dounce homogenizer and pestle type should I use. I've decided to change the cell lysis method, because freezing-thawing method doesn't work, and I can't prepare active lysate for my translation study since then. Can you give me any tips or protocols for HeLa cells Dounce homogenization and recomend some companies or homogenizers cat.no to this aim?
Thanks in advance
Lots of companies produce homogenisers. Basically there are two types - differentiated by the head on the pestle. The types are ball style head and cylinder style, where the end of the pestle looks like a ball or like a cylinder. The ball ones are typically used for retaining sub-cellular compartments such as nuclei or mitochondria, whereas the cylindrical ones will disrupt the sub-cellular fractions. There are also a range of different clearances between the mortar and pestle for different size tissues etc.
I have seen volumes up to 50ish mL. Sigma-aldrich and VWR both sell homogenisers.