Blocking solution mistake, please help! - (Jan/15/2013 )
This is a quick question, for my blocking step in western blotting I used 5% skim milk. Instead of dissolving the skim milk in 1X TBST, I made a mistake and dissolved it in 1X TBST containing 5% BSA (which I normally use to dilute my antibodies). I went on and incubated the membrane overnight with primary antibody. I can't stop thinking about, I am worried, will my mistake affect my result? Please help!
It should be fine, BSA is commonly used as a blocking reagent -that is its function in the primary incubation.
Great! that made me feel better about my work. I guess I will find today how my results turn out. Thanks