Protocol Online logo
Top : New Forum Archives (2009-): : Protein Expression and Purification

Protein purification for adherent mammalian cells - (Sep/03/2013 )

Hello everybody

hope u all are doing well


my Question:


To lyse, for adherent cells, after washing with cold PBS, then add RIPA buffer.

Which one come first? do we need to trypsinized the cells first to detach, and after that we add RIPA ?


I thought that we need to detach the cells first, is it true?


You can lyse the cells directly in the dish if you wish, and then perform some sort of protein quantitation (e.g. Bradford or BCA assays).


You can also lift cells with trypsin or by other detaching methods and then lyse either based on cell number per volume or in a defined volume and then perform quantitation as above.


ok moderator. thank you very much