I am looking for some ideas about this one. I use B actin as the normalizator in my western blot. i use different dilution factors of the B actin antibody. For some samples i use 500X and for others i use 1000X. I wonder when i normalize my samples and calculate the relative expression levels for the different protein targets should i take the different dilution factors of the B actin in account or should i pretend that they have the same dilution factor?