11 replies to this topic

[cell the truth]

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

[SuMi]

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

[bob1]

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?

[cell the truth]

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

Any detergent such a triton will kill the cells!

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:

You could try electroporating the cells to allow large molecules to permeate.

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

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

how have you determined that they aren't in the cell?

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.

[cell the truth]

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

[Jon Moulton]

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

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

[cell the truth]

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

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

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.

[Jon Moulton]

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

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

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

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.

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.

[The Ace]

Try Saponin

[bob1]

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

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

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

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

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.

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.

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.

[cell the truth]

As i promised, i will report now what we did and how it worked: we used saponine and it worked very nice after some starting troubles. we used a concentration of 15ug/ml and reached a sensitivity about 40-50 fold higher when treated with saponin compared to controls.

thx everybody for looking this thread and giving me advices, especially Jon Moulton :-), didn't see your signature...

[bob1]

Good work, sounds like things are going well.