I will do a cell culture, but I can not use trypsin-EDTA because it will lysate the membrane proteins and I'm interested in quantifying those proteins. Is there another way to take off the cells without losing protein and viability?
It really varies from cell line to cell line.
Sometimes, you can use the EDTA without the trypsin (called Versene). Sometimes piptting the media vigorously on the cells can also dislodge them.
It really varies from cell line to cell line.
Sometimes, you can use the EDTA without the trypsin (called Versene). Sometimes piptting the media vigorously on the cells can also dislodge them.
also i have used EDTA for this, it will depend on the cell type too,
I used at 1-2 mM EDTA/PBS.
https://www.researchgate.net/post/Trypsin-Free_Detachment_of_cells_using_EDTA
using cell scrapers lift the mono-layer is another option.
https://biology.stackexchange.com/questions/23253/alternatives-to-trypsin-for-cell-detachment
Best Wishes,
Hi Javier,
We had the same requirement for our adhesion studies where we needed the surface proteins intact and so couldn't use trypisn.
We used the Cell Dissociation solution Buffer (C5914 SIGMA)- it made a big difference in our results.. Hope it works for you..
All the best!
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