before the O/N labeling with cy dyes, 280 absorption was lets say 10 times 320 after removing the free dye the 280 to 320 ratio became 5 but 320 did not increase just that the sample got diluted as pd column result in 1 ml and before labeling it was 0.5 ml can i say the sample aggregated? unfortunantly aggregation index is not possible as i dont have 350 i might be able to plot the 320 and 400 absorption with a straight line then see the 350 correspond where but thats not accurate and my professor would definetly not accept it for my thesis