Which quantity of purified protein in WB? - (Jun/30/2008 )
I want to run a WB with a purified protein (no cell lysate) and do not know how much I should load. I would be really thankful for any (quick) suggestion!
LabS
-LabS-
You don't need to run much if the protein is purified but it depends on the antibody you are going to use. If you know that the antibody is clean and sensitive I'd only run 0.25-0.5ug. If the antibody isn't quite that good run about 1-2ug. This should be plenty! If you aren't sure about this antibody or have never used it, I would recommend you run multiple lanes with increasing amounts of protein. This will also let you determine what the limits of detection are with this specific antibody.
-rkay447-
QUOTE (rkay447 @ Jun 30 2008, 04:17 PM)
You don't need to run much if the protein is purified but it depends on the antibody you are going to use. If you know that the antibody is clean and sensitive I'd only run 0.25-0.5ug. If the antibody isn't quite that good run about 1-2ug. This should be plenty! If you aren't sure about this antibody or have never used it, I would recommend you run multiple lanes with increasing amounts of protein. This will also let you determine what the limits of detection are with this specific antibody.
Thanks :-)
-LabS-