Cell detachment: alternative to trypsine/EDTA? - (Jun/17/2009 )
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
ciroyo on Jun 17 2009, 11:15 AM said:
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
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.com/page.aspx?ID=11850)
PS: if you try it out, can you let us know how efficient this is?
dpo on Jun 17 2009, 04:50 PM said:
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.
