I am detecting an additional band aside from the expected VP1, VP2, and VP3 bands. This is AAV9, and the extra band is appearing at 100-110 Kda. Ideas?
So basically instead of 3 bands you are seeing 4 bands? Do you think it could be a related band or a non specific band? By looking at the streak I guess it could be a non specific band.
Yes, there should be three bands, instead of the four. The extra band is the first (highest MW species) band on the gel. I believe that this may be an impurity (such as an HCP) that was not removed during the AAV IDX purification. I wanted to see if anyone was familiar with this additional band.
It's very unlikely that you get such a strong defined band as a contaminant and nothing else (like the smear above) inbetween the three expected VP bands and this band. It could be an artifact of the gel system (dimerized or oligomerized VPs). Did you try with fresh DTT or b-ME in your SDS-PAGE loading buffer? Western Blot could show if it's VP or not.
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