I am confused about what I should dilute my primary antibody in. I have been using 5% skim milk in TBST when I dilute my primary antibody and secondary. But I know someone in a different lab uses just TBST to dilute their primary antibody with NO skim milk at all and it seems they are able to visualize their bands better that using the skim milk method.
I do block my membrane in 5% skim milk for one hour before incubating my primary antibody overnight, so I'm thinking that it isn't necessary to dilute my primary antibody in 5% skim milk.
Does anyone have suggestions?