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Blocking solution mistake, please help! - (Jan/15/2013 )

Hi all,

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!

Thanks all!


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 :)