Western blot of apoptotic cells
Posted 06 April 2011 - 08:03 AM
The procedure I normally use for western blot is as follows: I have the cells in 6-well plates (each well having equal numbers of cells) incubated with the compound of interest for a certain time. Then I wash the cells 2x with PBS buffer before adding lysis buffer and scraping the cells off.
The problem now is that I am testing a compound that is inducing apoptosis in cancer cell lines (MCF-7) in a concentration dependent way. The apoptotic cells (so especially the wells with higher concentration of the compound) will float away and if I wash them I loose them. I only analyse the attached cells not the floating cells. Therefore, I get a weaker signal for the wells with higher concentration of compound.
What is the proper way to retrieve all cells from the 6-well plates? Any advice is much appreciated!
Posted 06 April 2011 - 07:08 PM
Posted 07 April 2011 - 01:01 AM