I encounter a problem almost everytime I do a WB; my ladder seems to 'dilute' adjacent slots. I don't know why it is doing this. I always cast my gells following protocol, and 'empty' slots I fill with an ammount of 4x sample buffer comparable to the ammount of 4x SB in other slots.
Could the problem be that the ammount of protein that I load is too high? I always load ~40ng (20uL) of protein. The gells that I cast are 15% acrylamide, and I use a 10 slots comb.
The ladder that I use is bought from Fermentas (Thermoscientific), it's productnumber is 26616. I load 5uL of ladder each time I do a WB.
Has anybody else encountered this problem? Or does anybody know why the ladder is spoiling my results?
With best regards,