How to detach an antibody from cell surface
Posted 26 January 2010 - 03:56 AM
As I have allready asked in topic title, can you tell me a good way to detach antibodies (after treating my cells with them for a period of time) from the cell membrane. The reason for this is next: i need to see if the treatment has caused the change in antibody production by the cell itself, and in ICH, I need to detect only cell-derived antibodies and not the treatment antibodies, if you know what I mean (since the tretment and produced antibodies are human origine, my secondary antibody recognizes both of them, and that`s the main problem). I have tried with extensive washing with PBS, but that doesn`t help a lot. Do you think that the pronase or trypsin treatment woud help?
Posted 26 January 2010 - 11:46 PM
(that is: Best regards, in my language )
Posted 29 January 2010 - 04:39 AM
Posted 31 January 2010 - 02:32 PM
Can you try reduce the treatment Ab to F(ab)2, then use anti-Fc conjugate to detect produced Ab?