Problems with detection of protein expressed in yeast - (Jun/04/2014 )
recently I tried to detect my protein by western blot. The protein was transformed in the yeast strain (S.cerevisiae), and the transformed colonies were grown overnight in selective liquid medium. This was pelleted and lysed to load it on a gel. However when I try to detect the protein by western blot I only saw background.
Could it be that my protein wasn't properly expressed? Or that it formed inclusion bodies... (because I tried it a lot to detect the protein, even with different tags and also different protein extractions). What are the best solutions to this problem?
even if it was in inclusion bodies it should show up on the western blot if you just lyse and load with sds loading buffer (inclusion bodies should solubilize when denatured).
have you tried to detect the tag instead of the protein?