lysing cells with lysate buffers

[beachbum]

Hi,

 

I am trying to add a chemical to some yeast cells but I found out that this chemical can't penetrate through the yeast cell wall. Then I thought of adding a lysate buffer to lyse the cell, therefore the cell wall wouldn't be a problem. But does lysing the cells prevent it from functioning and being "normal". Thanks.

[Yggdrasil]

There are various compounds (for example, saponin) which can be used to permeablize cell membranes in order to perform procedures like immunofluorescence staining. These compounds do not completely lyse the cell, but the do make the cells permeable to large hydrophilic compounds. However, these chemicals will likely disrupt normal cell function so you wouldn't be able to grow the cells after permeablizing them.

[mattbimbo]

hi beachbum, what you need to do will depejnd on the chemical and the yeast you are interested in. 'spheroplasting' may be a method you will need to consider.

[Run]

Indeed lysing the cell will result in cell death. You need permeabilize the cell, this will also effect it functions, but is less radical then complete lysis. PBS/0.1% Triton X-100 is often used.

[Immunologist]

Actually, I think with a buffer, you would kill your cells. But if your protein (or molecule) is negatively charged, you could get it in by electroporation, just like in DNA transfection.