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Cell detachment: alternative to trypsine/EDTA?


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3 replies to this topic

#1 ciroyo

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Posted 17 June 2009 - 12:15 AM

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

#2 Michaelro

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Posted 17 June 2009 - 04:42 AM

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

#3 dpo

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Posted 17 June 2009 - 06:50 AM

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.


#4 ciroyo

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Posted 17 June 2009 - 07:35 AM

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. :lol:




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