Permeabilize Eukaryotic Membranes - (Nov/17/2009 )

Hi All,

we try to treat eukaryotic cells with some antibiotics. Now it seems like some of the antibiotics (aminoglycosides) aren't properly in the cells, so we are looking for some stuff to permeabilize the cells. Any good ideas? Links?

Thx lots for your help

I just use 0.2% Triton X-100 for 5 minutes to permeablise my cells. Works for me

Any detergent such a triton will kill the cells! You could try electroporating the cells to allow large molecules to permeate. However, the antibiotics should enter the cells fine... how have you determined that they aren't in the cell?

bob1 on Nov 18 2009, 12:45 AM said:

Thats our opinion too. But i think we try this, at very low concentrations... i'll report how it worked.

bob1 on Nov 18 2009, 12:45 AM said:

Risky cause of contamination. But if anything else doesnt work, maybe.

bob1 on Nov 18 2009, 12:45 AM said:

we worked with different concentrations and saw that it has some slight effects. also we saw that some of the antibiotics work fine and others won't.

actually we're trying saponine, but without the expected success.

Actually i heard from somebody of another lab that they use DMSO. Do you think thats a good idea? Any experience?

Another option: Endo-Porter, an endosomal release agent.

http://www.gene-tools.com/node/24

Jon Moulton on Nov 18 2009, 05:47 PM said:

Seems like you propose your own product... at least you were involved in generating figures for the presentation. is this a market place here?
We are looking for kind of cheap-solutions, not a time consuming and expensive one.

Thanks anyway to everybody who responded, i'll write you soon our results.

cell the truth on Nov 19 2009, 04:58 AM said:

I'm not hiding my affiliation, I included the company name in my signature. I offered an option, you needn't follow it. Good luck with your cytosolic delivery project.

Try Saponin

Jon Moulton on Nov 19 2009, 10:18 AM said:

If I may throw in my 2c again, Gene-tools does good reagents in my experience. Often for these sorts of things the market and price are there because of the difficulties of doing it... you may find in the long run it is easier to buy something like Jon suggested.

On a similar vein, you could try something like Lipofectamine or other lipid delivery mechanism... may not work unless they are similar charge to DNA/RNA.