Bands from the first detection appear at the second detection but with a differe - (Oct/27/2011 )

Hi all,

I have detected a protein at 63kDa and re-used the membrane to detect the loading control at 42 kDa .
But doing so, the first protein was detected again (which is normal as it is the same secondary antibody), my problem is that the pattern was different, I have bands where there was nothing previously.
So I don't know which of the detection is the "real" one.

Any suggestion about how I could know which is "real"?

Thanks

how specific is the antibody for your control?

if it is specific then try mixing the control antibody with that of the 63 kDa protein, they are far enough separated that you should be able to identify them.

or detect the control first then you protein of interest and see if you get the same pattern.