actin MW problem in western blot - (May/18/2009 )
hey all,
i am doing western blots and for my loading control (actin), i am consistently getting a specific band at 55-60 kd (the expected size is 42 kd). these are 293 cells that have been infected with adenovirus containing a gene of interest. im having trouble explaining why my size is off. any thoughts?
-medstudent1-
my actin also comes down to 50-60KDa...I bought from Sigma.
-Curtis-
Curtis on May 19 2009, 02:20 AM said:
my actin also comes down to 50-60KDa...I bought from Sigma.
interesting. mine is from sigma as well (monoclonal actin). any explanation as to what it is running higher? bound to another protein? post-translational modifications? what cell line are you using.
-medstudent1-
We use Abcam A16 actin, it runs at about 46 kDa and is very strong. The ab we had from before (either sigma or santa cruz?) was a goat and very dirty with quite a few non-specific bands.
-bob1-
This is good to know!
-medchemgirl-