western blotting loading control
Posted 21 June 2009 - 08:00 AM
Posted 21 June 2009 - 05:22 PM
Posted 21 June 2009 - 06:37 PM
I am purifying the cell membrane.It is clean membrane preparations in theory.So what should I choose as the internal protein for the loading control.Some friends adivised me the Galpha for the loading control.Is it ok?What else I can get?Thank you.
If you are doing whole cell lysis GAPDH or actin will be fine. If you are doing clean membrane preparations then neither will be fine.
Posted 22 June 2009 - 12:38 AM
I do not know what is the best loading control for WB of total cell membrane proteins. My experience, for example, I had a protein anchored in plasma membrane, so I isolated crude plasma membrane and as loading control I used Na/K ATPase a typical plasma membrane protein. May be, it is not typical loading control for plasma membrane (or total cell membrane generally) but it seemed to work in my experiments.
GAPDH and actin are not membrane proteins (GAPDH is cytosolic marker). At the web site of Abcam.com you could find helpful information.
Posted 22 June 2009 - 07:49 AM