I know that is not a transfer problem because when I use precasted gradient gel the transfer is complete. Once I use Homemade gel I have a loss in LW molecular proteins (see attachment). During the run I observe a different speed of migration between bio-rad ladder and our samples (the Migration front of samples moves faster). Also, as you can see in the attachment, the low part of the ladder in not well defined. Also, there are discrepancies between ladder and samples (a protein which is known to have 44/42 kd was detected between 50 and 75 kD).