Help with protein extraction! - WI-38 cells (Oct/05/2006 )
I would like to ask for your help. I'm working with human lung fibroblasts, WI-38 cells. I can't get good total protein extract from the cells. The protein concentartion is very very low,which is probably due to bad scrapping. The cells are growing in 75cm flask and they are at 70-80% confluency, so should be fine for good extraction. Is it possible that I will use Trypsin, to get the cell pallet?, no scrapping?. Thank you for help and time
I add a RIPA lysis buffer and then tape the plate to the shaker at top speed for 5 min. Scrapping isn't really even necessary as all the cells are off.