protein extraction from transwell inserts
Posted 06 March 2012 - 01:31 AM
I am trying to establish a co-culture system of lung epithelial cells and fibroblasts using transwells with pore diameter 1μΜ and 0.4μΜ. I wonder if there is a different protocol for lysis of the cells grown on inserts.
In the case of plain cell culture dishes i used to wash the cells twice with ice cold PBS and then extract the total proteins by scrapping using the appropriate lysis buffer. Can I use the scrapper on the membrane of the insert or there is a danger to damage it? Is it possible to loose any buffer or protein extract through the transwell pores?
I would appreciate any advice very much!
Posted 06 March 2012 - 11:57 AM
Posted 06 March 2012 - 02:36 PM