3 replies to this topic

[ciroyo]

hello,

i have a question related to the detachment of adherent cells. common is to use Trypsine/EDTA. but in my experiments i analyse cell surface glycans, coupled to proteoglycans and glycoproteins and by using both these molecules will released from the cell surface. beside i read something about influence of EDTA on hyaluronic acid, non-covalently coupled to proteoglycans, such as CD44. so i had lots of problems to detect hyaluronic acid at the cell surface of cells after detachment. i need, when possible, the most content of glycans after detachment of cells.

so my question is, is there another protocol about cell detachment beside trypsine/EDTA? i heard about citric saline...but is that a good alternative?

thanks a lot

ciroyo

[Michaelro]

ciroyo, on Jun 17 2009, 11:15 AM, said:

hello,

i have a question related to the detachment of adherent cells. common is to use Trypsine/EDTA. but in my experiments i analyse cell surface glycans, coupled to proteoglycans and glycoproteins and by using both these molecules will released from the cell surface. beside i read something about influence of EDTA on hyaluronic acid, non-covalently coupled to proteoglycans, such as CD44. so i had lots of problems to detect hyaluronic acid at the cell surface of cells after detachment. i need, when possible, the most content of glycans after detachment of cells.

so my question is, is there another protocol about cell detachment beside trypsine/EDTA? i heard about citric saline...but is that a good alternative?

thanks a lot

ciroyo

Hi
I'd recommend You to check about the cell dissociation solution from Sigma
I used it once but I don't know its formulation.
Maybe it will be helpful to You
Best
Michaelro

[dpo]

Nunc has some plates from which you can detach your cells by shifting the cells from 37 degrees to 20 degrees. I haven't tried these, but they might be ideal for what you. You can even order a free sample to try it out (http://www.nuncbrand...e.aspx?ID=11850)

PS: if you try it out, can you let us know how efficient this is?

Edited by dpo, 17 June 2009 - 06:51 AM.

[ciroyo]

dpo, on Jun 17 2009, 04:50 PM, said:

Nunc has some plates from which you can detach your cells by shifting the cells from 37 degrees to 20 degrees. I haven't tried these, but they might be ideal for what you. You can even order a free sample to try it out (http://www.nuncbrand...e.aspx?ID=11850)

PS: if you try it out, can you let us know how efficient this is?

thank you for this link. i ordered it directly. i write you about my experience with that dishes.