I am interested in (partial) permeabilization of plasma membranes of live cells in such a way which would ...
a) ... not lead to permeabilization of endocompartment's membranes (chiefly of ER membranes)
b) ... allow to permeation of small molecules through PM (in or out of the cytoplasm)
c) I would need method with the most replicable parameters across experiments (not for Immuno or visualisation - I intend to use it for some kinetics inside cells)
(d) - it is not necessary to be reversible, however it would be positive bonus :-) )
I have read some inspirational stories with saponins, I have heard of possibilities to use CHAPS or PEG. What are your real experiencies? What would You recommend?
(saponins are promising, but I feel headache from hard way to achieve replicability with them .. to even get properly defined saponins seems problem, working concentration? Solubility.. temperature dependent effect etc)
Most use digitonin for this purpose.
http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.367.5228&rep=rep1&type=pdf
Cheers
We have used PEG for permeabilization of plasma membranes of live plant (tobacco) protoplast cells (no cell wall) to take in DNA molecules (plasmids). Or they called PEG-mediated genetic transformation using protoplasts.
Electroporation is another way. Using electrical shock on the living cells such as bacteria. People are using this way for cells to take in DNA molecules for E. coli and other microbes. You need an electroporator, and the bacterial cells need to be 'competent' (or electro-grade competent).
Some people also use electroporation to deliver DNA into plant cells (protoplasts). I have not done this. But researchers have observed that the cells will die much more this way than using PEG-mediated method.
I know DNA can be delivered in, but I am not sure if they can reverse out.
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